БИОМАРКЕРЫ, ПРИГОДНЫЕ ПРИ ДИАГНОСТИКЕ ФИБРОЗА ПЕЧЕНИ

Настоящее изобретение относится к способам диагностики фиброза печени у субъекта, включающим определение уровней экспрессии плазминогена урокиназного типа, матричной металлопротеиназы 9 и β-2-микроглобулина, вычисление на их основании балльной оценки и постановку диагноза. Также настоящее изобретение относится к набору для диагностики фиброза печени, содержащему первое антитело, специфически связанное с активатором плазминогена урокиназного типа (uРА), второе антитело, специфически связанное с матричной металлопротеиназой 9 (ММР9), и третье антитело, специфически связанное с β-2-микроглобулином (β-2-MG). 6 н. и 11 з.п. ф-лы, 33 табл., 3 пр.

АЦЕТИЛ-КОФЕРМЕНТ А-КАРБОКСИЛАЗА 2 КАК МИШЕНЬ ДЛЯ РЕГУЛЯЦИИ СЖИГАНИЯ ЖИРА, НАКОПЛЕНИЯ ЖИРА, ЭНЕРГЕТИЧЕСЕОГО ГОМЕОСТАЗА И ДЕЙСТВИЯ ИНСУЛИНА

... 1. Способ стимуляции окисления жирных кислот и потери в весе у индивидуума, включающий стадию ингибирования активности ацетил-КоА-карбоксилазы 2 у указанного индивидуума. 2. Способ по п.1, где указанная активность ингибируется путем введения указанному индивидууму ингибитора ацетил-КоА-карбоксилазы 2 (АСС2). 3. Способ по п.1, где указанный индивидуум страдает патофизиологическим состоянием. 4. Способ по п.3, где указанное патофизиологическое состояние выбрано из группы, состоящей из ожирения и диабета. 5. Способ снижения уровней сахара в крови индивидуума, включающий стадию введения указанному индивидууму ингибитора ацетил-КоА-карбоксилазы 2 (АСС2). 6. Способ по п.5, где указанный индивидуум страдает диабетом. 7. Трансгенная мышь, геном которой содержит гомозиготный разрыв в эндогенном гене АСС2 кодирующем изоформу ацетил-КоА-карбоксилазу 2 ацетил-КоА-карбоксилазы, где указанный разрыв приводит к инактивации указанного гена, и где указанная мышь не продуцирует какой-либо функциональной ...

СПОСОБЫ И КОМПОЗИЦИИ ДЛЯ ПОЛУЧЕНИЯ ИНДУЦИРОВАННЫХ ГЕПАТОЦИТОВ

... 1. Способ получения индуцированного гепатоцита из негепатоцитной клетки, включающий предоставление негепатоцитной клетки, контактирование негепатоцитной клетки с агентом, который повышает или индуцирует экспрессию или активность одного или более факторов перепрограммирования, и культивирование негепатоцитной клетки в условиях, подходящих для перепрограммирования негепатоцитной клетки в индуцированный гепатоцит, посредством этого из негепатоцитной клетки получается индуцированный гепатоцит.2. Способ получения индуцированного гепатоцита из негепатоцитной клетки, включающий предоставление негепатоцитной клетки, введение в негепатоцитную клетку препарата на основе нуклеиновых кислот, которые кодируют, или препарата на основе белков, которые представляют собой один или более факторов перепрограммирования, и культивирование негепатоцитной клетки в условиях, пригодных для перепрограммирования негепатоцитной клетки в индуцированный гепатоцит, посредством этого из негепатоцитной клетки получается ...

High-content imaging of microfluidic devices

The present invention is related to high-content microscopy imaging of microfluidic cell culture systems. A method of high-content microfluidic device microscopy is contemplated, along with related statistical analysis and microfluidic device adaptors.

Cellular predictive models for steatosis

Methods for generating models for predicting biological activity of a stimulus on a test population of cells are provided. In particular computer-implemented methods for producing models for classifying a hepatocyte or population of hepatocytes according to whether it exhibits steatosis and also cholestasis or phospholipidosis are presented. Also models are produced for classifying stimuli based on hepatoxicity. The methods may involve receiving a set of phenotypic features of the cells or population of cells that have been exposed to stimuli and treated with one or more markers for particular cellular components by automated image analysis. A subset of the cell populations may be identified to be used in generating a model from data associated with the subset.

Cellular predictive models for toxicities

Normalizing cell assay data for models

Compositions and methods ofr determining anti-viral drug susceptibility and resistance and anti-viral drug screening.

This invention provides a method for determining susceptibility for an anti-viral drug comprising: (a)introducing a resistance test vector comprising a patient-derived segment and an indicator gene into a host cell; (b)cultring the host cell from (a); (c)measuring expression of the indicator gene in a target host cell; and (d)comparing the expression of the indicator gene from (c)with the expression of the indicator gene measured when steps (a)-(c)are carried out in the absence of the anti-viral drug, wherein a test concentration of the anti-viral drug is present at steps (a)-(c); at steps (b)-(c); or at step (c). This invention also provides a method determining anti-viral drug resistance in a patient. This also provides a method for evaluating the biological effectiveness of a candidate anti-viral drug compound. Compositions including resistance test vectors comprising a patient derived segment and an indicator gene and host cells transformed with the resistance test vectors are provided ...

PROCEDURE FOR MODULATING THE ACTIVITY OF TH2 CELLS BY MODULATING THE ACTIVITY OF XBP-1

LE-CELLS (LIVER ENGRAFTING CELLS), ASSAYS AND USES OF IT

PHOSPHORYLIERTE SP1 AS MARKER WITH THE DIAGNOSIS NICHTALKOHOLI STEATOHEPATITIS (NASH) AND A GOAL WITH DROGENSCREENING ON NASH

MULTI-WAVING MICRO EXAMINATION BY ABLATION

TEST PROCEDURE FOR CANDIDATE CONNECTIONS ON SUSCEPTIBILITY FOR BILIÄRE ELIMINATION

PROCEDURE AND COMPOSITIONS FOR MODULATING THE HEPATOZYTENWACHSTUMS, THE DIFFERENTIATION OF PLASMA CELLS OR THE ACTIVITY OF T-CELLS BY MODULATING THE ACTIVITY OF XBP-1

MOLECULES, ITSELF IN SELECTED ORGANS OR FABRICS THE INVIVO APPEARING AND PROCEEDING FOR YOUR IDENTIFICATION

USE OF STING AGONISTS TO TREAT CHRONIC HEPATITIS B VIRUS INFECTION

Provided herein are methods of treating a subject having hepatitis B viral (HBV) infection. More specifically, disclosed herein are methods of stimulating the innate cytokine response in macrophages, dendritic cells and/or liver non-parenchymal cells with small molecular agonists of STING to suppress HBV replication in hepatocytes. The methods are especially suitable for use in the treatment of chronic HBV infections. Also disclosed herein are methods of identifying compounds useful in the treatment of HBV infection.

Methods and compositions relating to modulation of hepatocyte growth, plasma cell differentiation of T cell subset activity by modulation

Cellular microarrays for screening differentiation factors

Provided is a microarray platform for the culture of cells atop combinatorial matrix mixtures; enabling the study of differentiation in response to a multitude of microenvironments in parallel.

Pharmaceutical compositions for and methods of inhibiting HCV replication

Highly functional liver cells derived from pluripotent stem cells, method for producing same, and method for testing metabolism/toxicity of drug

... [Problem] To produce functional liver cells, which are usable in testing the metabolism and toxicity of a drug, from pluripotent stem cells. [Solution] A method for producing highly functional liver cells using pluripotent stem cells, said method being characterized by comprising, from the pluripotent stem cells, acquiring primitive endoderm derived from the pluripotent stem cells by a process involving steps (A) and (B), from the primitive endoderm derived from the pluripotent stem cells, acquiring liver precursor cells by a process involving step (C), and, from the liver precursor cells, acquiring the highly functional liver cells by a proecss involving step (D): (A) a step for culturing under serum-free and feeder-free conditions; (B) a step for culturing in the presence of albumin and at least one kind of cytokine; (C) a step for culturing in the presence of SHH or an SHH agonist and at least one kind of cytokine; and (D) a step for culturing and maturing in the presence of at least ...

Method of screening candidate compounds for susceptibility to biliary excretion

A method of screening a candidate compound for susceptibility to biliary excretion by a hepatocyte transport protein. The method includes the steps of providing a culture of hepatocytes comprising a transport protein, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion by the transport protein. In some embodiments determining the amount of candidate compound in the bile canaliculus comprises inhibiting expression of the transport protein, measuring the amount of candidate compound in the bile canaliculus and comparing amounts of compound in the canalicules with and without inhibition of the transport protein. A difference in the amount of candidate compound in the canaliculus indicates susceptibility of the candidate ...

METHOD FOR DETERMINING THE INFLUENCE OF NON-PHYSIOLOGICAL COMPOUNDS THE DERIVATIVES AND DECOMPOSITION PRODUCTS THEREOF ON ORGANISMS ORGANS AND CELLS

Method for cell differentiation

Abstract The present invention relates to the field of cell biology, in in particular to methods for differentiating pluripotent stem cells. The invention provides methods for differentiating 5 primate pluripotent stem cells into cells of all three germinal layers. In particular, methods for differentiating primate pluripotent stem cells into the definitive endoderm are provided.

Utility of protein in the prediction of in vivo effects

A method of evaluating disposition and/or effect of a candidate compound in an in vitro culture and/or suspension to predict in vivo disposition and/or effect of the candidate compound, which can include the steps of providing a cell culture and/or suspension; exposing a candidate compound to the culture and/or suspension; exposing the culture and/or suspension to a media providing an in vivo relevant extracellular environment, such as a media comprising a component (such as a protein and/or other component, such as lipoproteins, bile acids, and endogenous compounds such as bilirubin) at a physiologic concentration or a concentration having characteristics, such as binding characteristics, similar to the physiological concentration; wherein any combination of any of the exposing steps can occur in any order or simultaneously; and evaluating in vitro disposition and/or effect of the compound to predict in vivo disposition and/or effect of the candidate compound.

ACETYL-COENZYME A CARBOXYLASE 2 AS A TARGET IN FAT REGULATION AND INSULIN ACTION

The present invention highlights the role of acetyl-CoA carboxylase through its product malonyl-CoA in regulating fatty acid oxidation and synthesis, glucose metabolism and energy homeostasis. It discloses transgenic mice with inactivating mutations in the endogenous gene for the acetyl-CoA carboxylase 2 isoform of acetyl-CoA caboxylase. Inactivation of acetyl-CoA caroxylase 2 results in mice exhibiting a phenotype of reduced malonyl-CoA levels in skeletal muscle and heart, unrestricted fat oxidation, and reduced fat accumulation in the liver and fat storage cells. As a result, the mice consume more food but accumulate less fat and remain leaner than wild-type mice fed the same diet. These results demonstrate that inhibition of ACC2 acetyl-CoA carboxylase could be used to regulate fat oxidation and accumulation for purposes of weight control. The instant invention provides a useful animal model to regulate malonyl-CoA production by ACC2 in the regulation of fatty acid oxidation by muscle ...

METHOD OF SCREENING CANDIDATE COMPOUNDS FOR SUSCEPTIBILITY TO BILIARY EXCRETION

A method of screening a candidate compound for susceptibility to biliary excretion. The method includes the steps of providing a culture of hepatocytes, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion. Optionally, the culture of hepatocytes is a long-term culture in a sandwich configuration. The method is particularly applicable to the screening of multiple candidate compounds in a single effort.

CELL VIABILITY DETECTION USING ELECTRICAL MEASUREMENTS

A method of determining information about cell viability and other characteristics relating to cell membrane permeability is disclosed. The method involves determining the effect of a cell on current flow and relating that effect to a known standard which standard may be a known healthy cell and thereby deducing the viability of the cell being tested. The cells being tested can be subjected to different environmental conditions such as surrounding chemicals, temperature, pH and pressure to determine the effects of such conditions on cell viability and/or cell permeability. The cell being tested can be in a cell suspension, grown on s ubstarte, in tissue in vitro or in tissue in vivo. The method provides substantially instantaneous results and need not include the use of dyes or other markers.

ISLET CELLS FROM HUMAN EMBRYONIC STEM CELLS

This disclosure provides a system for producing pancreatic islet cells from embryonic stem cells. Differentiation is initiated towards endoderm cells, and focused using reagents that promote emergence of islet precursors and mature insulin-secreting cells. High quality populations of islet cells can be produced in commercial quantities for use in research, drug screening, or regenerative medicine.

METHOD OF PRODUCING CELLS - PRECURSORS OF MATURE LIVER

IMPROVED PREPARATIONS CELLS - PRECURSORS OF MATURE LIVER

ВЕКТОРЫ ДЛЯ ОПРЕДЕЛЕНИЯ ЧУВСТВИТЕЛЬНОСТИ ВИРУСА К ПРОТИВОВИРУСНОМУ ЛЕКАРСТВЕННОМУ СОЕДИНЕНИЮ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

Настоящее изобретение относится к способу определения чувствительности вируса к противовирусному лекарственному препарату, включающему (а) культивирование клеток-хозяев в присутствии указанного противовирусного препарата, причем указанные клетки-хозяева содержат вектор для определения указанной чувствительности, содержащий фрагмент вируса и индикаторный ген, причем экспрессия указанного индикаторного гена зависит от активности гена, кодируемого указанным фрагментом вируса, и указанный вектор содержит не все вирусные гены, необходимые для образования инфекционной вирусной частицы; (б) измерение уровня экспрессии указанного индикаторного гена в указанных клетках-хозяевах; и (в) сравнение уровня экспрессии указанного индикаторного гена, измеренного на стадии (б), с контрольным уровнем экспрессии указанного индикаторного гена, причем разница между уровнем экспрессии указанного индикаторного гена, измеренным на стадии (б), и указанным контрольным уровнем экспрессии коррелирует с чувствительностью ...

COMPOSITIONS AND METHODS OF DETERMINING SENSITIVITY AND RESISTANCE TO CLAIMS A MEDICINAL AGENT AND SCREENING OF ANTIVIRAL DRUGS

Technical method for co-cultivating hepatocytes and hepatic sinusoidal endothelial cells and application of technical method

Selecting compounds that modulate reverse transport of cholesterol, useful e.g. for treating atherosclerosis, from their ability to bind to the liver receptor homolog-1 response element

La présente invention concerne des méthodes et composés susceptibles de moduler le transport inverse du cholestérol chez un mammifère ainsi que des méthodes de criblage permettant de sélectionner, d'identifier et/ou de caractériser des composés capables de moduler le transport inverse du cholestérol. Elle concerne également des cellules, vecteurs et constructions génétiques utilisables pour la mise en oeuvre de ces méthodes, ainsi que des compositions pharmaceutiques destinées au traitement de l'athérosclérose Les méthodes de l'invention sont basées sur l'utilisation d'éléments de réponse à LRH-1 dérivés du promoteur du gène apo AI.

위-장-간 생체 모사 마이크로 칩

... 위-장-간 생체 모사 마이크로 칩이 개시되며, 이러한 위-장-간 생체 모사 마이크로 칩은 분석대상물질이 위액과 함께 유입된 다음, 중화액에 의해 중화된 소화물 상태로 유출되는 위 채널, 및 장 세포가 배치되고 상기 중화된 소화물 중 적어도 일부가 상기 장 세포를 통과하도록 상기 위 채널과 연결되는 장 채널을 포함하는 위-장 채널부; 및 간 세포가 배치되고, 상기 장 세포를 통과한 물질 중 적어도 일부가 상기 간 세포와 접촉되도록 상기 장 채널과 연결되는 간 채널부를 포함하되, 상기 간 채널부는 상기 간 세포와 접촉된 물질 중 적어도 일부가 타겟 조직과 접촉되도록 형성된다.

DUAL-TRANSFECTED CELLS LINES AS IN VITRO SCREENING TOOLS FOR PHARMACEUTICAL COMPOUND PROFILING: A MODEL FOR HEPTOBILIARY ELIMINATION

The invention is referring to several dual-transfected cell lines expressing human NTCP (Na/taurocholate Cotransporting Protein; SLC10A1) together with human BSEP (Bile Salt Export Pump; ABCB11) or human MRP2 (Multidrug Resistance Protein; ABCC2) suitable as an in vitro tool for pharmaceutical compound profiling particularly as a model for hepatobiliary elimination. © KIPO & WIPO 2007 ...

METHOD FOR IMPROVING DRUG METABOLISM OF HEPATOCYTE

The present invention relates to a production method of high-function hepatocyte, which differentiates into hepatocyte from human pluripotent stem cells, and increases the expression and activity of drug metabolizing enzyme and the physiological function of the hepatocyte by repetitively treating xenobiotics on the differentiated hepatocyte. COPYRIGHT KIPO 2017 ...

COMPOSITION FOR INDUCING DIRECT CONVERSION OF SOMATIC CELL INTO HEPATIC STEM CELL, HEPATIC CELL, OR CHOLANGIOCYTE

The present invention relates to: a composition for inducing direct conversion of somatic cells into at least one selected from a group consisting of induced hepatic stem cell (iHSC), hepatic cells and cholangiocytes; a method of direct conversion of the somatic cells into at least one selected from a group consisting of iHSC, hepatic cells and cholangiocytes comprising a step for introducing the composition into the somatic cells; and a pharmaceutical composition for preventing or treating liver diseases comprising at least one selected from a group consisting of hepatic cells differentiation-induced from direct conversion-induced iHSC, and the cholangiocytes. COPYRIGHT KIPO 2018 ...

Method of screening nucleic acid based material targeting ITIH1 for treating disease related with hyperglycemia

METHODS RELATING TO MODULATION OF TH2 CELL SUBSET ACTIVITY BY MODULATION OF XBP-1 ACTIVITY

METHOD FOR MANUFACTURING CILIARY MARGIN STEM CELLS

CELLULAR PREDICTIVE MODELS FOR TOXICITIES

Methods for generating models for predicting biological activity of a stimulus test population of cells are provided. The models may be used to classify or predict the effect of stimuli on cells. In certain embodiments, the methods involve receiving data comprising values for dependent variables associated with stimuli; preparing a set of cell populations based on the data received; identifying a subset of the cell populations to be used in generating a model from data associated with the subset, wherein the model is provided to predict activity of a test population.

NORMALIZING CELL ASSAY DATA FOR MODELS

Methods for generating models for predicting biological activity of a stimulus test population of cells are provided. The models may be used to classify or predict the effect of stimuli on cells. In certain embodiments, the methods involve receiving data comprising values for dependent variables associated with stimuli as applied to cell populations; preparing a set of cell populations based on the data received; identifying a subset of the cell populations to be used in generating a model from data associated with the subset, wherein the model is provided to predict activity of a test population.

PROTOZOAN PROTEIN PERTAINING TO PLASMODIUM INFECTION IN LIVER STAGE

The present invention addresses the problem of identifying a protein essential for plasmodium in a liver-infection stage to proceed to the next stage, pro viding a test method for searching for a compound obstructing this prote in, and providing a compound for hindering infection with plasmodium by obstructing the protein. The objective is achieved by a method of testing a subst ance for hindering infection with plasmodium, the test method being characterized in that an evaluation is made of whether or not merozoite formation in a liver stage of infection with plasmodium is obstructed by restraining action of LS-specific protein 2 (LISP2) in which plasmodium is specifically ex pressed in host cells in the middle to the latter period of the liver stage.

IMAGING OF ENZYME ACTIVITY

This invention relates to biochemistry and magnetic resonance imaging.

Animal model for flaviviridae infection

The present invention is a woodchuck or an isolated woodchuck cell infected with bovine viral diarrhea virus. The invention can be used to identify new compounds for the treatment of flavivirus, pestivirus or hepatitis C infection using these models.

Compositions and methods for promoting lipid mobilization in humans

The invention provides methods of using polypeptide compounds based on the structures of insect peptides of the adipokinetic hormone family to mobilize lipids in humans. The compositions and methods described in the application are useful for modulating human body weight, such as inducing weight loss. The invention also includes screening methods for identifying other compounds effective for modulating lipid mobilization in humans.

COMPONENT ANALYSIS DEVICE, DRUG COMPONENT ANALYSIS DEVICE, COMPONENT ANALYSIS METHOD, AND DRUG COMPONENT ANALYSIS METHOD

Provided is a component analysis device, which comprises an analysis unit for measuring a component fed to the plurality of containers and analyzing the component thus measured, wherein the plurality of containers are at least a first container and a second container, wherein the first container retains a first solution containing a substance that releases adhesion between multiple cells forming the liver cell tissue, and the second container retains a buffer solution, and wherein the analysis unit measures an amount of the component discharged from the liver cell tissue in the first container into the first solution and an amount of the component discharged from the liver cell tissue in the second container into the buffer solution in the second container, and analyzes an amount of the component to be discharged via a bile duct in the liver cell tissue. 1. A component analysis device , comprisinga retention unit for retaining a plurality of containers for retaining a liver cell tissue, andan analysis unit for measuring a component fed to the plurality of containers and analyzing the component thus measured, whereinthe plurality of containers are at least a first container and a second container, whereinthe first container retains a first solution containing a substance that releases adhesion between multiple cells forming the liver cell tissue, andthe second container retains a buffer solution, and whereinthe analysis unit measuresan amount of the component discharged from the liver cell tissue in the first container into the first solution andan amount of the component discharged from the liver cell tissue in the second container into the buffer solution in the second container, and analyzesan amount of the component to be discharged via a bile duct in the liver cell tissue.2. The component analysis device according to claim 1 , further comprising a temperature regulation unit for regulating a temperature of a liquid in the plurality of containers 1 , whereinthe ...

Method for high-throughput screening of compounds and combinations of compounds for discovery and quantification of actions, particularly unanticipated therapeutic or toxic actions, in biological systems

The invention enables high-throughput screening of compounds in living systems to detect unanticipated or unintended biological actions. The invention also allows for screening, detection, and confirmation of new indications for approved drugs. Screening and detection of toxic effects of compounds also can be achieved by using the methods of the invention. The methods comprise administering isotope-labeled substrates to a living system so that the label is incorporated into molecules in a manner that reveals flux rates through metabolic pathways thought to be involved in a disease. Comparisons between living systems exposed to compounds and living systems not so exposed reveals the effects of the compounds on the flux rates through the metabolic pathways. Combinations or mixtures of compounds can be systematically screened to detect unanticipated or unintended biological actions, including synergistic actions, in the same manner.

Molecules that home to a selected organ or tissue in vivo and methods of identifying same

The present invention provides an in vivo method for identifying molecules that home to a selected organ or tissue. In addition, the invention provides peptides that home to a selected organ or tissue. For example, the invention provides peptides that selectively home to an organ such as brain or kidney or to a tissue such as a tumor tissue. The invention further provides methods of using an organ homing molecules, for example, to target an agent such as a drug to a selected organ or to identify the target molecule expressed by the selected organ.

肝実質細胞中のＰ４５０タンパク質のアイソフォームの質量分析による定量

ГЕНЕТИЧЕСКИ КОДИРУЕМЫЕ ИНДИКАТОРЫ ИОНОВ КАЛИЯ

Настоящее изобретение относится к области биотехнологии, конкретно к пептидным калиевым сенсорам, и может быть использовано для детекции положительно заряженных ионов калия. Конструируют полипептид, содержащий по меньшей мере один сигнальный домен и калиевый сенсор, который может связываться с K. При этом первый сигнальный домен может генерировать детектируемый сигнал после связывания Kс калиевым сенсором. Изобретение обеспечивает эффективную детекцию Kв образце. 7 н. и 18 з.п. ф-лы, 7 табл., 12 пр., 7 ил.

КОМПОЗИЦИЯ КУЛЬТУРАЛЬНОЙ СРЕДЫ И СПОСОБ КУЛЬТИВИРОВАНИЯ КЛЕТКИ ИЛИ ТКАНИ С ИСПОЛЬЗОВАНИЕМ УКАЗАННОЙ КОМПОЗИЦИИ

КОМПОЗИЦИЯ КУЛЬТУРАЛЬНОЙ СРЕДЫ И СПОСОБ КУЛЬТИВИРОВАНИЯ КЛЕТКИ ИЛИ ТКАНИ С ИСПОЛЬЗОВАНИЕМ УКАЗАННОЙ КОМПОЗИЦИИ

... 1. Композиция среды, способная культивировать клетки или ткани путем суспендирования их, которая имеет вязкость не более 8 мПа·с (в условиях 37˚C) и содержит деацилированную геллановую камедь или ее соль.2. Композиция среды по п. 1, где концентрация деацилированной геллановой камеди или ее соли в композиции среды составляет 0,01-0,05% (масса/объем).3. Композиция среды по п. 1, дополнительно содержащая полисахарид, отличный от деацилированной геллановой камеди или ее соли.4. Способ получения композиции среды по п. 1, который включает смешение деацилированной геллановой камеди или ее соли и среды.5. Способ получения композиции среды по п. 4, где среда является жидкой средой.6. Способ получения композиции среды по п. 4 или 5, который включает смешение деацилированной геллановой камеди или ее соли, растворенных или диспергированных в растворителе, и среды.7. Способ получения композиции среды по п. 6, где деацилированная геллановая камедь или ее соль, смешанная с растворителем или диспергированная ...

Probenpräparation für proteomische Untersuchungen

Es wird ein Verfahren zum Aufschluss einer Probe biologischen Materials zur Proteomanalyse durch ein massenspektrometrisches Verfahren bereitgestellt, wobei die Probe mit einem bestimmten Volumen einer organischen Säure versetzt, für einen bestimmten Zeitraum zur Lyse inkubiert und anschließend neutralisiert wird.

Cellular Predictive Models For Toxicities

Novel diagnostic and therapeutic method

An autotaxin(ATX)-lysophosphatidic acid(LPA) signalling pathway modulator for use in treating hepatitis B infection. The modulator may be an antagonist of a G-protein coupled lysophosphatidic acid receptor (LPAR), an inhibitor of hypoxia inducible factor 1 alpha (HIF1α) such as a phosphoinositide 3 kinase (PI3K) or mitogen activated protein kinase (MAPK) or MAPK/MEK inhibitor. Specifically the modulator may be HA130, siRNA targeting of ATX or HIF1α, KM-6425, wortmannin, BYL-719 or TGX-221.

Method and reagents for treating hepatic fibrosis and inflammation

Islet cells prepared from stem cells

The invention discloses a method for producing pancreatic islet cells from embryonic stem cells, such as primate pluripotent stem cells. The islet cells are characterised in that they produce at least one of insulin, glucagons, somatostatin or a pancreatic polypeptide Differentiation is promoted to wards endoderm cells using islet cell differentiation factors including nicotinamide. Also claimed are methods of producing insulin, screening methods for modulators and devices for generating such islet cells. Pharmaceutical preparations and methods of treatment, especially of type 1 diabetes are claimed.

Novel toxicity assay based on human blastocyst-derived stem cells and progenitor cells

Cell lines expressing p450 enzymes

Cultured cell lines are transfected with adenovirus expression vectors that encode at least one functional cytochrome P450 enzyme, wherein the cytochrome P450 (CYP) gene or genes are expressed in tandem as bi-cistronic or poly-cistronic fusions, separated by self-processing cleavage sequences, such as the 2A sequence from FMDV (foot-and-mouth virus). More than one CYP gene may be expressed in this way, and/or a CYP gene may be co-expressed with a cytochrome P450 reductase (CPR) gene, particularly where the cell lacks endogenous CPR activity. A reporter gene such as luciferase or epitope-tagged -hCG under the control of a cellular stress response element (eg the antioxidant response element - ARE) may also be present on the vector and linked to a self-processing cleavage sequence. The cell lines may be used for screening compounds and therapeutic agents for toxicity. The cells are preferably liver cells.

TOXIZITÄTSSCREENINGVERFAHREN

HEPATITIS C VIRUs TEST SYSTEMS

IMMORTALISIERTE HEPATOZYTEN

ISOLATED LIVER MAIN CELLS

HEPATOBLASTEN AND PROCEDURES FOR YOUR ISOLATION

PROTEIN-INDUCING MORPHOGENESE

METHOD OF MEASURING HUMAN CYP3A INDUCIBILITY

Hcv antiviral and cytotoxicity drug screening assay

Insulin-independent, bone morphogenetic protein (BMP)-mediated uptake of blood glucose by peripheral cells and tissues

Three dimensional cell patterned bioploymer scaffolds and method of making the same

Protein-induced morphogenesis

Compositions and methods for determining anti-viral drug susceptibility and resistance and anti-viral drug screening

PROTEIN-INDUCED MORPHOGENESIS

Islet cells from human embryonic stem cells

Biomarkers for the treatment of hepatocellular carcinoma

Use of STING agonists to treat chronic hepatitis B virus infection

Provided herein are methods of treating a subject having hepatitis B viral (HBV) infection. More specifically, disclosed herein are methods of stimulating the innate cytokine response in macrophages, dendritic cells and/or liver non-parenchymal cells with small molecular agonists of STING to suppress HBV replication in hepatocytes. The methods are especially suitable for use in the treatment of chronic HBV infections. Also disclosed herein are methods of identifying compounds useful in the treatment of HBV infection.

INSULIN-INDEPENDENT, BONE MORPHOGENETIC PROTEIN (BMP)-MEDIATED UPTAKE OF BLOOD GLUCOSE BY PERIPHERAL CELLS AND TISSUES

HUMAN HEPATIC PROGENITOR CELLS AND METHODS OF USE THEREOF

Liver progenitor cells immunoreactive for CD117, as well as for CD34 capable of proliferating in a culture; and differentiating in vivo into a hepatocyte, a cholangiocyte or a sinusoidal cell are provided. The cultures can be expended over a large number of passages and integrate well after transplantation into adult liver.

METHOD OF SCREENING CANDIDATE COMPOUNDS FOR SUSCEPTIBILITY TO BILIARY EXCRETION

HUMAN HEPATIC PROGENITOR CELLS AND METHODS OF USE THEREOF

... ²²²Liver progenitor cells immunoreactive for CD117, as well as for CD34 capable ²of proliferating in a culture; and differentiating in vivo into a hepatocyte, ²a cholangiocyte or a sinusoidal cell are provided. The cultures can be ²expended over a large number of passages and integrate well after ²transplantation into adult liver.² ...

HIGH-CONTENT IMAGING OF MICROFLUIDIC DEVICES

The present invention is related to high-content microscopy imaging of microfluidic cell culture systems. A method of high-content microfluidic device microscopy is contemplated, along with related statistical analysis and microfluidic device adaptors.

LIVER ENGRAFTING CELLS, ASSAYS, AND USES THEREOF

A substantially enriched mammalian hepatic liver engrafting cell population is provided. Methods are provided for the isolation and culture of this liver engrafting cell. The progenitor cells are obtained from a variety of sources, including fetal and adult tissues. The cells are useful in transplantation, for experimental evaluation, and as a source of lineage and cell specific products, including mRNA species useful in identifying genes specifically expressed in these cells, and as targets for the discovery of factors or molecules that can affect them.

METHOD OF MEASURING HUMAN CYP3A INDUCIBILITY

A method of measuring human CYP3A inducibility at the administration of a test drug characterized by comprising, to a non-human animal to which the test drug has been administered or cultured human cells having been cultured in a medium containing the test drug, transferring (A) a virus constructed by implanting a detectable reporter gene and at least 3 sites of regions capable of binding to human PXR in the non-translation region of human CYP3A gene into an adenovirus vector, and (B) a virus constructed by implanting human PXR gene into an adenovirus vector, and then measuring the expression dose of the reporter gene in the non-human animal or the cultured human cells. According to this method, the human CYP3A inducibility at the administration of a test drug to humans can be conveniently and accurately evaluated, which makes it possible to accurately evaluate the efficacy of the test drug, the expression of its side effect, the disappearance of the drug effect, etc.

COMPOSITIONS AND METHODS FOR INDUCTION OF PROTEINS INVOLVED IN XENOBIOTIC METABOLISM

The present invention provides improved cells and methods for identifying compounds that alter protein expression, such as xenobiotics, endobiotics, chemical or drugs. One aspect of the present invention is a cell that includes a first nucleic acid molecule that includes: a promoter or enhancer operable for a nucleic acid molecule encoding a protein involved in drug metabolism (such as an enzyme or transporter) and a reporter gene; and a second nucleic acid encoding an intracellular receptor or transcription factor; so that when the intracellular receptor or transcription factor is bound with a compound, the intracellular receptor or transcription factor can operably bind with the promoter or enhancer resulting in the expression of said reporter gene. Another aspect of the present invention is a method for evaluating compounds for the property of inducing the expression of a gene encoding a protein involved in drug metabolism, including; providing a test compound; contacting the test compound ...

MODEL SYSTEM OF LIVER FIBROSIS AND METHOD OF MAKING AND USING THE SAME

Provided herein is a model system for liver fibrosis, including a liver extracellular matrix, and a combination of mammalian liver cells (e.g., primary liver cells) on the matrix. In some embodiments, the combination of liver cells includes: (a) liver progenitor cells, (b) Kupffer cells, and (c) hepatic stellate cells. Methods of making the model system and methods of use of the model system for screening active agents are also provided.

HEPATOCYTE PRODUCTION VIA FORWARD PROGRAMMING BY COMBINED GENETIC AND CHEMICAL ENGINEERING

The present invention provides methods comprising both genetic and chemical means for the production of hepatocytes from a variety of cell sources, particularly pluripotent stem cells.

BIOMARKERS FOR THE TREATMENT OF HEPATOCELLULAR CARCINOMA

Provided herein are biomarkers for hepatocellular carcinoma and uses thereof.

CULTURE MEDIUM COMPOSITION, AND METHOD FOR CULTURING CELL OR TISSUE USING SAID COMPOSITION

The present invention provides: a method for culturing a cell and/or a tissue, said method being characterized by culturing the cell and/or the tissue in a floated state using a culture medium composition, wherein amorphous structures are formed in a liquid culture medium, are dispersed in the solution uniformly, and substantially hold the cell and/or the tissue without substantially increasing the viscosity of the solution, so that the culture medium composition has an effect of preventing the sedimentation of the structures; and others.

METHOD TO STUDY INSULIN ACTION

This invention relates to a method of studying insulin action in a cell line, wherein the cell line is an immortalised cell line. This method comprises contacting the cell line with a factor, and determining whether the factor affects or mimics insulin action.

MEDIUM FOR CELL CULTURE MLEKOPITAYuShchIKhSYa, METHOD OF ITS PRODUCTION AND MIXTURE OF COMPONENTS, COMPOSITION, METHODS FOR PREPARING, PRODUCING FOR CLONAL GROWTH, REPRODUCTION OF AND CHANGING PHENOTYPE OF HEPATOCYTES IN VITRO OF, CELLS, PHARMACEUTICAL COMPOSITION, METHODS FOR PREPARING RECOMBINANT CELLS AND GENE PRODUCT, METHODS OF USING CELLS, METHOD FOR TRANSPLANTATION OF HEPATOCYTES, METHOD OF TESTING A MEDICINAL PREPARATION, METHOD FOR MAINTAINING VIABILITY OF DIFFERENTIATED HEPATOCYTES

Pharmaceutical composition for suppression of formation of kzkdnk hepatitis b virus

COMPOSITION AND KIT COMPRISING OF PIPERAZINE DERIVATIVES AND METFORMIN, THEIR USE IN THE TREATMENT OF DIABETES

B형 간염 바이러스의 ｃｃｃＤＮＡ 형성 억제용 약학 조성물

... 본 발명은 B형 간염 바이러스 항체가 B형 간염 바이러스의 표면 항원(HBsAg)이 헤파란 황산 프로테오글라이칸(heparan sulfate proteoglycan)에 결합하는 것을 방해함으로써 B형 간염 바이러스의 cccDNA의 형성을 억제한다는 것을 확인한 것에 기초한 것이다. 본 발명의 약학 조성물 및 B형 간염 바이러스의 cccDNA 형성 억제 방법을 활용하면 만성 B형 간염을 근본적으로 치료할 수 있고 B형 간염 환자의 간이식 수술 후 간염 재발을 예방할 수 있을 것으로 기대된다. 또한, 본 발명에 따르면 cccDNA 형성 억제 여부 확인을 통해 B형 간염 예방 또는 치료용 물질을 새로이 동정(identify)할 수 있다. 또한, 본 발명에 따르면, B형 간염 바이러스 항체와 병용투여 될 수 있는 치료 보조제를 새로이 동정(identify)할 수 있다.

모양체 주연부 간세포의 제조 방법

... 본 발명은, 이하의 공정 (1) 혹은 공정 (2)의 어느 하나 또는 이들 모두의 공정을 포함하는, 다능성 간세포로부터 분화 유도된 모양체 주연부 간세포의 제조 방법을 제공한다: (1) 다능성 간세포로부터 분화 유도된 모양체 주연부 유사 구조체를 포함하는 세포 응집체로부터 얻어진 세포를 부유 배양해, 레티노스피어를 얻는 공정；및 (2) 다능성 간세포로부터 분화 유도된 모양체 주연부 유사 구조체를 포함하는 세포 응집체로부터 얻어진 세포로부터 단계 특이적 배성 항원-1 양성 세포를 분취하는 공정. 본 발명에 의해, 망막층 특이적 신경세포를 포함하는 망막 세포로의 분화능 및 자기 복제능을 갖는 모양체 주연부 간세포를 고순도로 효율적으로 제조하는 것이 가능해진다.

세포 배양 방법 및 스크리닝 방법

... 생체 내 기능을 장기간 유지할 수 있으며 또한, 필요 최소 세포수에서의 배양이 가능한 세포 배양 방법을 제공하는 것을 목적으로 한다. 본 발명에 관련된 세포 배양 방법은 구획된 미소 공간 내에 있어서, 미분화 세포를 중층화한 상태에서 배양하여, 분화 세포를 얻는 것이다. 의약품을 스크리닝하는 경우, 간 세포, 장 상피 세포, 신경 세포, 심근 세포, 혈관 내피 세포로 분화되는 미분화 세포가 바람직하다. 특히, 약물 동태 등의 인간에 대한 예측을 실시하는 경우, 인간 세포인 것이 바람직하다.

Infected cell cultures

CULTURE MEDIUM COMPOSITION, AND METHOD FOR CULTURING CELL OR TISSUE USING SAID COMPOSITION

METHODS FOR PRODUCING CELLS HAVING A PHENOTYPE OF A PRIMARY HUMAN HEPATOCYTES AND COMPOSITIONS

Methods of predicting responsiveness to interferon treatment and treating hepatitis c infection

Provided are methods of predicting responsiveness of a subject to interferon treatment, comprising comparing a level of expression in a cell of the subject of at least one gene selected from the group consisting KIR3DL3, KIR3DL2, KIR3DL1, KIR2DL1, KIR2DL2, KIR2DL3, KLRG1, KIR3DS1, CD160, HLA-A, HLA-B, HLA-C, HLA-F, HLA-G and IFI27 to a reference expression data of the at least one gene obtained from at least one interferon responder subject and/or at least one interferon non-responder subject. Also provided are methods and pharmaceutical compositions for treating a subject in need of interferon treatment.

Use of inhibitors of scavenger receptor class proteins for the treatment of infectious diseases

The present invention relates to the use of inhibitors of scavenger receptor class proteins, in particular ScarB1 for the production of a medicament for treatment of and/or prophylaxis against infections, involving liver cells and/or hematopoietic cells, in particular malaria.

Methods of Modulating Thrombocytopenia and Modified Transgenic Pigs

The application provides methods of modulating platelet uptake by liver sinusoidal endothelial cells and of modulating thrombocytopenia. Transgenic pigs modified to bind fewer platelets are provided.

HEPATOCYTE PRODUCTION BY FORWARD PROGRAMMING

The invention generally features methods for providing hepatocytes from a variety of cell sources, particularly pluripotent stem cells, therapeutic compositions featuring such cells, and methods of using them for the treatment of subjects. 1. A method of producing hepatocytes by forward programming of stem cells , comprising: culturing the stem cells under conditions to artificially increase the expression level of FOXA2 , HNF4A and one or more additional hepatocyte programming factor genes selected from the group consisting of HHEX , HNF1A , FOXA1 , TBX3-1 , GATA4 , NR0B2 , SCML1 , CEBPB , HLF , HLX , NR1H3 , NR1H4 , NR1I2 , NR1I3 , NR5A2 , SEBOX , ZNF391 , and ZNF517 , thereby producing hepatocytes from forward programming of the stem cells.2. The method of claim 1 , wherein the stem cells comprise at least one exogenous expression cassette comprising one or more of the hepatocyte programming factor genes.3. The method of claim 1 , comprising contacting the stem cells with hepatocyte programming factors comprising gene products of the hepatocyte programming factor genes in an amount sufficient to cause forward programming of the stem cells to hepatocytes.4. The method of claim 3 , wherein the gene products are polypeptides or RNA transcripts of the hepatocyte programming factor genes.5. The method of claim 1 , wherein the stem cells are mesenchymal stem cells claim 1 , hematopoietic stem cells claim 1 , or induced pluripotent stem cells.6. The method of claim 1 , wherein the number of the hepatocyte programming factor genes are two claim 1 , three claim 1 , four claim 1 , five or six.7. The method of claim 1 , wherein the hepatocyte programming factor genes comprise Forkhead box protein A1 (FOXA1) claim 1 , forkhead box A2 (FOXA2) claim 1 , hematopoietically-expressed homeobox protein (HHEX) claim 1 , hepatocyte nuclear factor 1 homeobox A (HNF1A) claim 1 , hepatocyte nuclear factor 4 alpha (HNF4A) claim 1 , GATA binding protein 4 (GATA4) claim 1 , NR0B2 (nuclear ...

IN VITRO MODEL FOR PATHOLOGICAL OR PHYSIOLOGIC CONDITIONS

The present invention generally relates to in vitro methods for mimicking in vivo pathological or physiologic conditions. The methods comprise applying shear forces to a cell type or cell type plated on a surface within a cell culture container. Methods for testing drugs or compounds in such systems are also described. 1. A method of mimicking a pathological or physiologic condition in vitro , the method comprising:adding a culture media to a cell culture container;adding at least one factor to the culture media;plating at least one cell type on at least one surface within the cell culture container; andapplying a shear force upon the at least one plated cell type, the shear force resulting from flow of the culture media induced by a flow device, the flow mimicking flow to which the at least one cell type is exposed in vivo in the pathological or physiologic condition, wherein: (i) within the in vivo concentration range of the factor observed in the pathological condition; or', '(ii) within the concentration range of the factor that would result in vivo from administration of a drug or a compound; or, 'the concentration of the factor in the culture media for mimicking the pathological condition is either (i) within the in vivo concentration range of the factor observed in the physiologic condition; or', '(ii) within the concentration range of the factor that would result in vivo from administration of a drug or a compound., 'the concentration of the factor in the culture media for mimicking the physiologic condition is either2. An in vitro method of testing a drug or a compound for an effect on the pathological or physiologic condition , the method comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'mimicking the pathological or physiologic condition according to the method of ;'}adding a drug or a compound to the culture media; andapplying the shear force upon the at least one plated cell type exposed to the drug or the compound; wherein a change in the ...

HEPATOCYTES FOR IN VITRO GENOTOXICITY TESTS

The invention relates to a method for carrying out genotoxicity tests of chemical, biological and physical active substances or agents with the aid of cell culture systems of proliferating physiologically active liver cells. 1. A method of testing genotoxicity in vitro , the method comprising using proliferating hepatocytes for carrying out the in vitro genotoxicity testing.2. The method according to claim 1 , further comprising carrying out the testing using the Ames test claim 1 , chromosome aberration test claim 1 , comet assay and micronucleus test.3. The method according to claim 1 , wherein the proliferating hepatocytes comprise at least four Phase I enzymes selected from the group consisting of CYP1A2 claim 1 , CYP2C9 claim 1 , CYP2C19 claim 1 , CYP2D6 claim 1 , CYP2E1 claim 1 , CYP3A4 claim 1 , CYP1A2 claim 1 , CYP2A6 claim 1 , CYP2B6 claim 1 , CYP2C8 claim 1 , CYP1A1 claim 1 , CYP3A5 claim 1 , CYP3A7 and CYP4A11.4. The method according to claim 1 , wherein the proliferating hepatocytes exhibit no viability in soft agar or no tumor growth in vivo.5. The method according to claim 1 , wherein the proliferating hepatocytes are from primary cells of humans or mammals and comprise at least one ofa) a proliferation gene, andb) at least one cellular factor which is inactivated that induces cell division arrest, andc) are transiently immortalized.6. The method according to claim 5 , wherein the cellular proliferation gene is selected from the group consisting of at least one of myc claim 5 , jun claim 5 , ras claim 5 , src claim 5 , fyg claim 5 , myb claim 5 , E2F claim 5 , Mdm2 claim 5 , TERT; an E6 or E7 proliferation gene of papillomaviruses; a large or small tumor antigen of polyomaviruses; E1A or E1B protein of adenoviruses; an EBNA protein of the Epstein Barr virus; HTLV and Herpesvirus saimiri.7. The method according to claim 5 , wherein the viral proliferation gene is selected from the group consisting of at least one of E6 and E7 proliferating genes of HPV ...

Method to Identify Liver Toxicity Using Metabolite Profiles

The present disclosure relates to a methodology that identifies the underlying mechanisms that lead to hepatotoxicity. This is done by using the alterations in cellular metabolite profiles obtained before and after therapy/drug treatment in combination with a model of liver metabolism. Subsequently, by using a covariance matrix adaptation followed by evolutionary selection, it compares the drug-induced metabolite profiles obtained experimentally with those generated using the in silico model, automatically shortlists potential parameters whose alterations could have produced the drug-treated metabolite profile. Values of these parameters are estimated by formulating an optimal control problem to minimize the differences between the model-generated metabolite values and experimentally observed data. The estimated parameters are given as an input to the homeostatic liver model and simulations are carried out, thereby the results providing a mechanistic explanation for the development of toxicity.

METHODS AND COMPOSITIONS FOR TREATING OBESITY AND/OR DIABETES AND FOR IDENTIFYING CANDIDATE TREATMENT AGENTS

Provided are methods and compositions for identifying candidate agents for treatment of obesity, liver disease, and/or diabetes. Such methods include, e.g., contacting a mammalian cell or cell population with a test agent, and measuring an expression level and/or activity level of ClpP in the mammalian cell or in cells of the cell population. Also provided are methods and compositions for treating an individual (e.g., one who is obese and/or has diabetes). Treatment methods include administering an inhibitor of ClpP to the individual (e.g., to prevent or reduce weight gain, to increase insulin sensitivity, and/or to increase glucose tolerance). 1. A method of identifying a candidate agent for treating obesity , liver disease , and/or diabetes , the method comprising:(a) contacting a mammalian cell with a test agent;(b) measuring an expression level and/or activity level of ClpP in the mammalian cell relative to a reference value following the contacting;(c) determining that the test agent caused a decrease in the expression level and/or activity level relative to the reference value; and(d) identifying the test agent as a candidate agent for treating obesity, liver disease, and/or diabetes.2. The method according to claim 1 , wherein the mammalian cell is a mouse cell.3. The method according to claim 1 , wherein the mammalian cell is a human cell.4. The method according to any one of - claim 1 , wherein the mammalian cell is in vitro.5. The method according to any one of - claim 1 , wherein the mammalian cell is ex vivo.6. The method according to any one of - claim 1 , wherein the mammalian cell is in vivo.7. The method according to any one of - claim 1 , wherein the mammalian cell is a hepatocyte.8. The method according to claim 2 , wherein the contacting comprises administering the test agent to a mouse.9. The method according to claim 8 , further comprising claim 8 , measuring an expression level and/or activity level of ClpP in the mouse prior to the contacting ...

Induced Hepatocytes and Uses Thereof

Disclosed herein are induced hepatocytes from a trophoblast stem cell, methods for inducing the cells, and compositions thereof. Also disclosed herein are methods of treating a disease or disorder (e.g., liver-associated) by utilizing an induced hepatocyte disclosed herein. 189-. (canceled)90. An isolated human hepatocyte , wherein the isolated human hepatocyte expresses human leukocyte antigen G (HLA-G) and human cytoplasmic marker stem 121 (stem 121).91. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocyte is a human hepatic progenitor cell.92. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocyte further expresses one or more biomarkers selected from the group consisting of CXCR4 claim 90 , FOXA2 claim 90 , SOX17 claim 90 , HHEX claim 90 , TTR claim 90 , ALB claim 90 , TAT claim 90 , CYP7A1 claim 90 , BSEP claim 90 , SERPINA1 claim 90 , G6PC claim 90 , ABCC2 claim 90 , C/EBPβ claim 90 , HNF1α claim 90 , HNF4α claim 90 , and any combination thereof.93. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocyte further expresses TGFβ1 claim 90 , fibronectin claim 90 , or collagen IV in extracellular matrix (ECM).94. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocyte recruits CD4Foxp3 Treg cells.95. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocytes form tissue of a 3-dimensional structure.96. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocytes cluster or aggregate.97. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocytes form a crescent cell mass.98. The isolated human hepatocyte of claim 90 , wherein the isolated human hepatocyte further expresses one or more markers selected from the group consisting of TGFβ1 claim 90 , C-kit claim 90 , CK19 claim 90 , CK18 claim 90 , ALB claim 90 , α-AFP claim 90 , betatrophin claim 90 , ADH1 claim 90 , APOF claim 90 , CPS1 claim ...

CELL CULTURE

Aspects of the present invention relate to apparatus for use in cell and tissue culture techniques. Particularly, although not exclusively, embodiments of the present invention relate to apparatus which contribute to providing a dynamic cell culture environment. Also disclosed herein are methods for culturing cells and/or tissues, together with in vitro methods of testing drug efficacy as well as other subject matter. 1. An apparatus for providing bi-directional fluid flow , the apparatus comprising:a culture apparatus for culturing cells and/or a tissue or portion thereof, and the culture apparatus comprising a holder body, the holder body comprising a plurality of chambers for containing a tissue culture media and at least one fluid communication pathway extending between at least two respective chambers of the plurality of chambers, each fluid communication pathway permitting bi-directional fluid flow between said at least two chambers, and wherein said apparatus for providing bi-directional fluid flow further comprises:a holder body support; andat least one drive element arranged to rock a holder body supported via the support to thereby repeatedly raise and lower spaced apart ends of the holder body, wherein the apparatus for providing bi-directional fluid flow is configured to repeatedly rock the holder body.2. The apparatus according to claim 1 , wherein each chamber of said plurality of chambers comprises a base element and at least one side wall element.3. The apparatus according to claim 2 , wherein:the fluid communication pathway comprises a through channel extending between a base element or side wall element of a first one of the plurality of chambers to a base element or side wall element of a further one of the plurality of chambers; or(b) the fluid communication pathway comprises a through slot extending between a base element or side wall element of a first one of the plurality of chambers to a base element or side wall element of a further one of ...

Methods and Systems for High-throughput Toxicity Screening of a Compound Using Mahalanobis Values

The invention relates to methods and systems for high-throughput toxicity screening of compounds using Mahalanobis Values, and in particular comparing a normal unexposed transcriptome Mahalanobis Value in an in-vitro hepatocyte microassay against a calculated transcriptome Mahalanobis Value of hepatocytes exposed to a target compound for varying time periods and in varying concentrations. 1. A method for displaying toxicity of a chemical , comprising the steps:Performing a transcriptome array analysis of a hepatocyte cell line exposed to the chemical, wherein the hepatocyte cell line is exposed to at least five different concentrations of the chemical ranging from 0.001-10 micromolar to form at least five concentration samples of the hepatocyte cell line, and each concentration sample of the five different concentration samples are exposed to the chemical for at least five different time periods ranging from 2-72 hours to form at least 25 concentration-duration samples of the hepatocyte cell line;Calculating an Abnormal Mahalanobis Distance Value and a Abnormal p-value for each concentration-duration sample of the at least 25 concentration-duration samples of the hepatocyte cell line;Calculating an Mahalanobis Distance number for 1-risk deviation, 2-risk deviations, and 3-risk deviations from a Normal Mahalanobis Distance, where the Normal Mahalanobis Distance is calculated from a transcriptome array analysis of a hepatocyte cell line unexposed to the chemical;Visually presenting the Abnormal Mahalanobis Distance Values on a grid of chemical concentrations against times of exposure, where the chemical concentrations are the at least five different concentrations of the chemical ranging from 0.001-10 micromolar and the times of exposure are the at least five different time periods ranging from 2-72 hours; and, Visually marking the grid of Abnormal Mahalanobis Distance Values to identify the 1-risk deviation, 2-risk deviations, and 3-risk deviations from Normal ...

Biomarkers useful in liver fibrosis diagnosis

Identification of urokinase-type plasminogen, matrix metalloproteinase 9, and β-2-microglobulin as novel biomarkers associated with liver fibrosis and uses thereof in diagnosing and staging liver fibrosis.

IN-VITRO METHODS FOR CLASSIFYING DRUGS ACCORDING TO THEIR POTENTIAL TO CAUSE LIVER CELL INJURY

The invention provides in vitro methods for classifying drugs according to their potential to cause liver injury 1. An in vitro method of classifying a drug according to the drug's potential to cause liver cell injury , comprising:a) obtaining a population of liver cells;b) contacting the population of liver cells with a drug provided at a range of concentrations;c) contacting the population of liver cells with a cytokine provided at a range of concentrations;d) determining cytotoxicity of the population of liver cells;e) generating a concentration-response curve;f) defining covariates from the curve using a four-parameter logistic model;g) developing a classification model using logistic regression of covariates defined in step h) to generate a logistic regression model; andi) evaluating by receiver operating characteristic (ROC) the optimal classification model and covariate combination, to thereby classify the drug according to the drug's potential to cause liver injury.2. The method of claim 1 , wherein the liver cells are primary human heptocytes.3. The method of claim 1 , wherein the hepatoma cells are human hepatoma cells.4. The method of claim 1 , wherein the human hepatoma cells are HepG2.5. The method of claim 1 , wherein the liver cell injury is a hepatocellular injury.6. The method of claim 1 , wherein the liver cell injury is liver death.7. The method of claim 5 , wherein the hepatocellular injury is idiosyncratic claim 5 , drug-induced liver injury (IDILI).8. The method of claim 1 , wherein the drug is selected from the group consisting of steroidal or nonsteroidal anti-inflammatory drugs (NSAIDs) claim 1 , antibiotic claim 1 , anti-viral claim 1 , anti-bacterial claim 1 , anti-fungal claim 1 , chemotherapeutic claim 1 , small molecule drugs of any pharmacologic class claim 1 , cardiac claim 1 , pulmonary claim 1 , lipid-modulating claim 1 , neuromodulatory claim 1 , analgesic claim 1 , drugs that modify blood coagulation claim 1 , gastrointestinal (GI ...

CELL-BASED BIOIDENTITY TEST FOR INSULIN

A functional cell-based assay for use as a bioidentity assay for insulin or insulin analogs is described. The assay may be used as a replacement of the rabbit blood sugar method disclosed in USP<121> Insulin Assays. 1. A method for determining the potency of an insulin or insulin analog in a sample comprising:(a) providing a cell culture of recombinant cells capable of gluconeogenesis, wherein the recombinant cells comprise a nucleic acid molecule encoding a reporter molecule operably linked to the promoter for the glucose-6-phosphate catalytic-subunit-encoding gene C (G6PC);(b) mixing equal aliquots of the cell culture with a series of dilutions of cell culture medium comprising the insulin or insulin analog to provide a series of assay cell cultures and incubating the series of assay cell cultures for about 15 to 27 hours;(c) detecting the reporter molecule in the series of assay cell cultures to provide data for the assay cell culture and generating a dose response curve from the data; and (i) mixing equal aliquots of the cell culture with a series of dilutions of a reference cell culture medium comprising a known amount of the insulin or insulin analog to provide a series of reference assay cell cultures,', '(ii) incubating the series of reference assay cell cultures for about 15 to 27 hours, and', '(iii) detecting the reporter molecule in the series of reference assay cell cultures to provide data for the reference assay cell culture and generating the reference dose response curve from the data, to provide the potency of the insulin or insulin analog in the sample., '(d) comparing the dose response curve to a reference dose response curve obtained from one or more series of reference assay cell cultures determined by a method comprising'}2. (canceled)3. The method of claim 1 , wherein the recombinant cells are H4IIE rat hepatoma cells or HepG2 human hepatoma cells claim 1 , which comprise the nucleic acid molecule encoding the reporter molecule operably linked ...

METHOD FOR PRODUCING HEPATIC STEM/PRECURSOR CELLS FROM MATURE HEPATIC CELLS USING LOW-MOLECULAR-WEIGHT COMPOUND

The present invention provides a method for producing hepatic stem/progenitor cells from mammal hepatocytes, comprising bringing a TGFβ-receptor inhibitor, and optionally a GSK3 inhibitor and/or a ROCK inhibitor into contact with the hepatocytes in vitro. 1. A method for producing hepatic stem/progenitor cells from mammal hepatocytes , comprising bringing a TGFβ-receptor inhibitor into contact with the hepatocytes in vitro.2. The method according to claim 1 , further comprising bringing a GSK3 inhibitor and/or a ROCK inhibitor into contact with the hepatocytes in vitro.3. The method according to claim 1 , further comprising bringing a GSK3 inhibitor and a ROCK inhibitor into contact with the hepatocytes in vitro.4. The method according to claim 1 , wherein the contact between the hepatocytes and the TGFβ-receptor inhibitor is carried out by culturing the hepatocytes in the presence of the inhibitor.5. The method according to claim 2 , wherein the contact between the hepatocytes and the GSK3 inhibitor and/or the ROCK inhibitor is carried out by culturing the hepatocytes in the presence of the inhibitor.6. The method according to claim 1 , wherein the mammal is a human claim 1 , a rat or a mouse.7. An agent for inducing hepatic stem/progenitor cells from hepatocytes claim 1 , which comprises a TGFβ-receptor inhibitor.8. The agent according to claim 7 , which is combined with a GSK3 inhibitor and/or a ROCK inhibitor.9. The agent according to claim 7 , which is combined with a GSK3 inhibitor and a ROCK inhibitor.10. The agent according to claim 7 , wherein the hepatocytes are derived from a human claim 7 , a rat or a mouse.11. Hepatic stem/progenitor cells derived from mammal hepatocytes claim 7 , comprising the following characteristics:(a) have self-regeneration ability;(b) capable of differentiating into both hepatocytes and biliary epithelial cells; and(c) express EpCAM but not Dlk1 as a surface antigen marker.12. An agent for inducing hepatic stem/progenitor cells ...

COMPONENT ANALYSIS DEVICE, DRUG COMPONENT ANALYSIS DEVICE, COMPONENT ANALYSIS METHOD, AND DRUG COMPONENT ANALYSIS METHOD

Provided is a component analysis device, which comprises an analysis unit for measuring a component fed to the plurality of containers and analyzing the component thus measured, wherein the plurality of containers are at least a first container and a second container, wherein the first container retains a first solution containing a substance that releases adhesion between multiple cells forming the liver cell tissue, and the second container retains a buffer solution, and wherein the analysis unit measures an amount of the component discharged from the liver cell tissue in the first container into the first solution and an amount of the component discharged from the liver cell tissue in the second container into the buffer solution in the second container, and analyzes an amount of the component to be discharged via a bile duct in the liver cell tissue. 1. A component analysis device , comprisinga retention unit for retaining a plurality of containers for retaining a liver cell tissue, andan analysis unit for measuring a component fed to the plurality of containers and analyzing the component thus measured,wherein the plurality of containers are at least a first container and a second container,wherein the first container retains a first solution containing a substance that releases adhesion between multiple cells forming the liver cell tissue,wherein the second container retains a buffer solution, andwherein the analysis unit:measures an amount of the component discharged from the liver cell tissue in the first container into the first solution and an amount of the component discharged from the liver cell tissue in the second container into the buffer solution in the second container, andanalyzes an amount of the component to be discharged via a bile duct in the liver cell tissue.2. The component analysis device according to claim 1 , further comprising a temperature regulation unit for regulating a temperature of a liquid in the plurality of containers claim 1 , ...

COMPOSITION FOR PREVENTING OR TREATING LIVER FIBROSIS OR CIRRHOSIS, COMPRISING EXPRESSION OR ACTIVITY ENHANCER OF TIF1y AS ACTIVE INGREDIENT

The present invention relates to a composition for preventing and treating liver fibrosis or cirrhosis and, more specifically, to a pharmaceutical composition for preventing and treating liver fibrosis or cirrhosis, comprising an expression or activity enhancer of transcriptional intermediary factor 1 gamma (TIF1γ) as an active ingredient, and a method for screening the same. The pharmaceutical composition for preventing or treating liver fibrosis or cirrhosis, comprising an expression or activity enhancer of TIF1γ as an active ingredient, according to the present invention, inhibits the activity of hepatic stellate cells (HSCs) and decreases the expression of α-SMA proteins or the secretion of collagen Type I, thereby ultimately being expected to be developed as a prophylactic or therapeutic agent for liver fibrosis or cirrhosis. In addition, the composition of the present invention is expected to be useful in a method for screening an agent for liver fibrosis or cirrhosis. 15.-. (canceled)6. A method for screening a candidate material for preventing or treating liver fibrosis or cirrhosis , comprising steps of (1) treating cells or tissues harvested from a patient with liver fibrosis or cirrhosis with a test material and culturing the treated cells or tissues; (2) measuring an expression level of TIF1γ in a cell or tissue culture solution of Step (1); and (3) selecting a candidate material which increases the expression of TIF1γ as compared to a control which is not treated with the test material.7. The method of claim 6 , wherein the test material is a synthetic compound claim 6 , a microbial culture solution or extract claim 6 , a synthetic peptide claim 6 , a nucleic acid claim 6 , a protein claim 6 , an antibody claim 6 , an aptamer claim 6 , or a natural extract.8. A method for preventing or treating liver fibrosis or cirrhosis claim 6 , comprising administrating to a subject claim 6 , a pharmaceutical composition comprising an expression enhancer or activity ...

Lipopeptides for Use in Treating Liver Diseases and Cardiovascular Diseases

The present invention relates to lipopeptide-based compounds for use in the diagnosis, prevention and/or treatment of a liver disease or condition, preferably liver involved metabolic diseases, as well as in the control or modification of the cholesterol level or cholesterol uptake and, thus, diagnosis, prevention and/or treatment of a cardiovascular disease. The present invention furthermore relates to an in vitro or in vivo assay or method for testing or measuring the NTCP-mediated transport of test compound(s). The present invention furthermore relates to a method for the diagnosis, prevention and/or treatment of a liver disease or condition, comprising administering a therapeutically effective amount of a lipopeptide-based compound to a patient. The present invention furthermore relates to a method for the diagnosis, prevention and/or treatment of a cardiovascular disease.

Methods for administering pexidartinib

The present disclosure relates generally to methods for using pexidartinib and related Risk Evaluation and Mitigation Strategy (REMS), while reducing the occurrence of hepatotoxic adverse events.

HIGH DENSITY 3D HEPATOCYTE SPHEROID PLATFORM FOR DRUG ADME STUDIES

The present disclosure relates to methods for evaluating the interaction of a candidate compound on 3D hepatocyte spheroid in an invitro culture, including evaluating the metabolism of a candidate compound, for use in various biochemical and molecular biology studies. The methods are performed in labware that combine 3D spheroid culture with micro-patterned design that allows for multiple to several hundreds of spheroids to be treated under the same conditions and to produce sufficient materials (e.g., parent drug, drug metabolites, DNA, RNA, and proteins from cells) and higher detection signal intensity for ADME/Tox (absorption, distribution, metabolism, excretion and toxicity) studies. The methods allow for, among other uses, the investigation and generation of accurate invitro intrinsic clearance data and thus more accurate prediction of in vivo clearance, particularly with low clearance compounds. 1. An assay method for evaluating the interaction of one or more low clearance candidate compounds with hepatocytes , comprising: 'a top aperture; and a liquid impermeable bottom comprising a bottom surface, wherein at least a portion of the bottom comprises a low-adhesion or no-adhesion material in or on the bottom surface;', 'culturing hepatocytes in a cell culture article to form a spheroid, wherein the cell culture article comprises a chamber, the chamber comprising an array of microcavities, each microcavity structured to constrain the hepatocytes to grow in a 3D spheroid confirmation to form a hepatocyte spheroid; wherein each microcavity of the chamber comprisescontacting the 3D hepatocyte spheroid with one or more low clearance candidate compounds; andmeasuring the in vitro intrinsic clearance of the one or more candidate compounds.2. The assay method of claim 1 , wherein the culture is a long-term culture.3. (canceled)4. The assay method of claim 1 , wherein the liquid impermeable bottom comprising the bottom surface is gas-permeable.5. The assay method of ...

CULTURE MEDIUM COMPOSITION AND METHOD OF CULTURING CELL OR TISSUE USING THEREOF

The present invention provides a culture method of cells and/or tissues including culturing cells and/or tissues in a suspended state by using a medium composition wherein indeterminate structures are formed in a liquid medium, the structures are uniformly dispersed in the solution and substantially retain the cells and/or tissues without substantially increasing the viscosity of the solution, thus affording an effect of preventing sedimentation thereof, and the like. 1. A medium composition comprising a structure capable of culturing cells or tissues by suspending them.2. The medium composition according to claim 1 , permitting an exchange treatment of the medium composition during culture and recovery of the cells or tissues after completion of the culture.3. The medium composition according to claim 1 , which does not require any of a temperature change claim 1 , a chemical treatment claim 1 , an enzyme treatment and a shear force claim 1 , during recovery of the cells or tissues from the medium composition.4. The medium composition according to claim 1 , having a viscosity of not more than 8 mPa·s.5. The medium composition according to claim 1 , wherein the aforementioned structure has a size that passes a filter having a pore size of 0.2 μm to 200 μm when it is passed through a filter.6. The medium composition according to claim 1 , wherein the aforementioned structure contains a polymer compound.7. The medium composition according to claim 6 , wherein the aforementioned polymer compound includes a polymer compound having an anionic functional group.8. The medium composition according to claim 6 , wherein the aforementioned polymer compound is a polysaccharide.9. The medium composition according to claim 7 , wherein the aforementioned anionic functional group is at least one kind selected from the group consisting of a carboxy group claim 7 , a sulfo group and a phosphate group.10. The medium composition according to claim 8 , wherein the aforementioned ...

INCREASED AQUAPORIN EXPRESSION ON CELLULAR MEMBRANE TO IMPROVE CRYOPRESERVATION EFFICIENCY

A method of storing mammalian cells or tissue (e.g., liver cells or hepatocytes) for subsequent use comprises the steps of: (a) contacting the cells or tissue in vitro to a choleretic agent in an effective amount; (b) combining said cells or tissue with a cryopreservative; (c) freezing said cells or tissue, and then (d) storing said frozen cells or tissue in frozen form for subsequent use. 1. A method of storing mammalian cells or tissue for subsequent use , comprising the steps of:(a) contacting the cells or tissue in vitro to a choleretic agent in an effective amount;(b) combining said cells or tissue with a cryopreservative; and then(c) freezing said cells or tissue.2. The method of claim 1 , further comprising the step of:(d) storing said frozen cells or tissue in frozen form for subsequent use.3. The method of claim 1 , wherein said cells or tissue are liver cells or tissue.4. The method of claim 1 , wherein said cells or tissue comprise hepatocytes.5. The method of claim 1 , wherein said choleretic agent comprises DiButyryl cAMP (BtcAMP) or glucagon.6. The method of claim 1 , wherein said cryopreservative comprises glycerol.7. The method of claim 1 , wherein said mammalian cells or tissue are human cells or tissue.8. The method of claim 1 , wherein said choleretic agent is contacted to said cells or tissue in an amount effective to increase the expression of aquaporins on cell membranes thereof.9. The method of claim 8 , wherein said aquaporins comprise Aquaporin-8 (AQP8).10. The method of claim 2 , wherein said storing step is carried out for at least one week.11. Frozen mammalian cells or tissues produced by the process of and stored in sterile form in a container.12. A method of providing live mammalian cells or tissue claim 1 , comprising the step of:{'claim-ref': {'@idref': 'CLM-00011', 'claim 11'}, '(a) thawing cells or tissue of .'}13. The method of claim 12 , further comprising the step of:(b) rinsing said cells or tissue with an aqueous solution to ...

METHOD OF PREPARING CELLS FOR 3D TISSUE CULTURE

The present invention relates to a method of preparing cells for 3D tissue culture, which method comprises the steps of plating the cells on a suitable surface, optionally, checking for their capability to adhere to said surface, discarding the cells which have not adhered to said surface, detaching the adhered cells and transferring them into a 3D tissue culture process. 1. A method of preparing cells for 3D tissue culture , which method comprises the steps ofa) plating the cells on a suitable surfaceb) optionally, checking for their capability to adhere to said surface,c) discarding the cells which have not adhered to said surface,d) detaching the adhered cells and transferring them into a 3D tissue culture process.2. The method of claim 1 , which does not encompass the application of a viability assay throughout steps a)-d).3. The method of claim 1 , wherein the 3D tissue culture process is at least one selected from the group consisting ofscaffold-based 3D tissue culturehydrogel-based 3D tissue culturecellular self-assembly 3D tissue culture, and/or3D bioprinting,4. The method of claim 1 , wherein the cells are cryopreserved cells which are thawed before plating.5. The method of claim 1 , wherein the cells are non-frozen cells.6. The method of claim 1 , wherein the cells are selected fromprimary cellsstem cells,tumor cells and/orimmortalized cells.7. The method of claim 1 , wherein the cells are not expanded prior to claim 1 , during or after plating claim 1 , and/or the cells are not passaged after plating.8. The method of claim 1 , wherein the cells are mammalian cells claim 1 , preferably selected from the group consisting ofhuman cellscynomolgus cellspig cellscanine cells,rat cells,9. The method of claim 1 , wherein the cells are Hepatocytes.10. A 3D tissue culture process selected from the group consisting ofscaffold-based 3D tissue culturehydrogel-based 3D tissue culturecellular self-assembly 3D tissue culture, and/or3D bioprinting,{'claim-ref': {'@idref': ...

APPLICATION OF ECM1 IN PREVENTION AND/OR TREATMENT OF LIVER FIBROSIS-RELATED DISEASES

Provided is application of ECM1 in the prevention and/or treatment of liver fibrosis-related diseases, specifically provided is the use of ECM1 gene, or protein or a promoter thereof for preparing a composition or a formulation, the composition or formulation being used for (a) preventing and/or treating of liver fibrosis-related diseases; and/or for (b) maintaining liver homeostasis. The ECM1 gene, or the protein or promoter thereof can significantly (i) prevent and/or treat cirrhosis-related diseases; and/or (ii) maintain the liver homeostasis. In addition, the ECM1 gene, or the protein or promoter thereof can also significantly (i) inhibit the occurrence of liver fibrosis-related diseases; and/or (ii) inhibit the activation of hepatic stellate cells (HSCs). 112-. (canceled)13. A method for (a) preventing and/or treating a liver fibrosis-related disease; and/or (b) maintaining liver homeostasis , the method comprising administering to a subject a pharmaceutical composition comprising an ECM1 gene , or a protein thereof or a promoter thereof , wherein the promoter comprises a substance that increases the expression of the ECM1 gene or increases the or activity of the ECM1 protein.14. The method of wherein the pharmaceutical composition inhibits the occurrence of a liver fibrosis-related disease; and/or (ii) inhibits the activation of a hepatic stellate cell (HSC).15. The method of wherein the pharmaceutical composition comprises an ECM1 promoter selected from the group consisting of a small molecule compound claim 13 , a vector expressing ECM1 claim 13 , and a combination thereof.16. The method of wherein the pharmaceutical composition comprises an ECM1 protein selected from the group consisting of:(A) A polypeptide whose amino acid sequence is shown in either SEQ ID NO: 1 or SEQ ID NO: 3;(B) an ECM1 protein derivative or an active fragment thereof formed by substitution, deletion or addition of one or more amino acid residues in the amino acid sequence as shown in ...

MASS SPECTROMETRY QUANTITATION OF P450 PROTEIN ISOFORMS IN HEPATOCYTES

A method for screening a drug for cytochrome P450 (CYP) induction is provided and can include incubating the drug with a microsome-containing biological sample and then quantitating at least one cytochrome P450 isoform. The isoforms can be selected from 2B6, 3A4, 1A2, and 3A5 isoforms. In some embodiments, the method uses liquid chromatography tandem mass spectrometry (LC-MSMS). A quantitated value can be compared to a threshold value and the drug can be determined to exhibit an acceptable CYP induction potential when the quantitated value does not exceed the threshold value. Isolated peptides are also provided. 1. A method for determining an amount of at least one isoform of cytochrome P450 (CYP) in a sample , said method comprising subjecting said sample to an analysis with a liquid chromatograph tandem mass spectrometer system , said system comprising a triple quadrupole instrument and Multiple Reaction Monitoring (MRM) , said analysis comprising detecting at least one isolated peptide of CYP 3A4 , CYP 1A2 or CYP 3A5.2. The method of wherein the inform is CYP 3A4 and said isolated peptide has the amino acid sequence of SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 claim 1 , or SEQ ID NO: 73. The method of claim 2 , wherein the amino acid sequence is SEQ ID NO:5 and said analysis is performed by monitoring a precursor-product ion pair transition having an m/z value of about 440/549 claim 2 , 440/650 claim 2 , or 440/532 in said system and processing data from said monitoring to determine said amount.4. The method of claim 2 , wherein the amino acid sequence is SEQ ID NO:6 and said analysis is performed by monitoring a precursor-product ion pair transition having an m/z value of about 704/794 claim 2 , 704/929 claim 2 , 564/689 claim 2 , 564/745 claim 2 , or 564/790 in said system and processing data from said monitoring to determine said amount.5. The method of claim 2 , wherein the amino acid sequence is SEQ ID NO:7 and said analysis is performed by monitoring a precursor- ...

METHOD FOR MEASURING BILE SALT EXPORT AND/OR FORMATION ACTIVITY

A method is provided to measure modulation of bile salt export transport and/or formation activity in hepatocyte or stable cell line preparations by test agents including but not limited to drugs, drug candidates, biologicals, food components, herb or plant components, proteins, peptides, DNA, RNA. Furthermore, the method is to determine modulation of bile salt export transport and/or formation activity not only by said test agents, but further their metabolites or biotransformed products formed in situ. The bile salt export transport and/or formation activity modulation includes but not limited to inhibition, induction, activation and/or regulation. The method can be practiced to identify test agents, which have potential to cause liver injury, drug-drug interactions, and/or can be used as therapeutic agents for the treatment of cholestasis, abnormality of bile salt metabolism, liver diseases and cholesterol abnormality. 1. A kit for assessing a test agent's effect on bile salt export transport and/or formation activity comprising one or more bile salt precursor compounds and one or more hepatocyte preparations , wherein the hepatocyte preparations are used to prepare hepatocyte suspensions for assessing a test agent's effect on bile salt export transport and/or formation activity.2. The kit of wherein the hepatocyte preparation comprises primary hepatocytes and/or hepatocytes derived from stable cell lines.3. The kit of wherein the hepatocyte preparation comprises hepatocytes derived from human or animal tissue.4. The kit of wherein the hepatocyte preparation comprises hepatocytes derived from human liver tissue.5. The kit of wherein the hepatocytes preparation comprises hepatocytes derived from mouse claim 3 , rat claim 3 , dog claim 3 , rabbit or monkey liver tissue.6. The kit of wherein the hepatocyte preparation comprises hepatocytes derived from stable cell lines.7. The kit of wherein the hepatocyte preparation comprises pooled hepatocytes derived from one or ...

IN VITRO MODEL FOR PATHOLOGICAL OR PHYSIOLOGIC CONDITIONS

The present invention generally relates to in vitro methods for mimicking in vivo pathological or physiologic conditions. The methods comprise applying shear forces to a cell type or cell type plated on a surface within a cell culture container. Methods for testing drugs or compounds in such systems are also described. 1299-. (canceled)300. A method for mimicking a pathological condition of the liver in vitro , the method comprising:adding a culture medium and at least one factor to a cell culture container;plating hepatocytes on at least one surface within the cell culture container, wherein the hepatocytes comprise hepatocytes isolated from at least one subject having the pathological condition; andapplying a shear force upon the plated hepatocytes, the shear force resulting from flow of the culture medium induced by a flow device, the flow mimicking flow to which the hepatocytes are exposed in vivo in the pathological condition.301. The method of claim 300 , wherein the hepatocytes comprise primary cells.302. The method of claim 300 , wherein the surface within the cell culture container comprises a porous membrane suspended in the cell culture container.303. The method of claim 300 , wherein the shear force is applied indirectly to the plated hepatocytes.304. The method of claim 303 , wherein the hepatocytes are plated on a first surface of a porous membrane claim 303 , the porous membrane is suspended in the cell culture container such that the first surface is proximal and in spaced relation to a bottom surface of the cell culture container claim 303 , thereby defining within the cell culture container a lower volume comprising the hepatocytes and an upper volume comprising a second surface of the porous membrane claim 303 , and the shear force is applied upon the second surface of the porous membrane in the upper volume of the container.305. The method of claim 304 , further comprising plating nonparenchymal hepatic cells on the second surface of the porous ...

IMPROVED PREPARATIONS OF ADULT LIVER PROGENITOR CELLS

Preparations of adult liver progenitor cells (called HHALPCs) have been manufactured from different human donors and characterized by using cell surface markers that allow identifying HHALPCs preparations and/or the methods for producing them that are most suitable for cell therapy, in particular for treating liver diseases or inherited blood coagulation disorders. 1. An isolated human adult liver progenitor cell characterized in that said cell is measured positive for:(a) the mesenchymal or pluripotent markers CD13, CD73, CD90, and CD105;(b) the adhesion markers CD29, CD44, CD47, CD49b, CD49c, CD49e, and CD147;(c) the tetraspanins CD9, CD63, CD81, and CD151; and(d) CD98, CD140b, and β2-microglobulin.2. The cell of characterized in that the cell is measured positive for:(a) at least one marker selected from adhesion markers CD54, CD164, CD165, and CD166; and/or(b) at least one marker selected from CD46, CD55, CD59, and CD95.3. The cell of characterized in that the cell is measured positive for at least one marker selected from CD26 claim 1 , CD49a claim 1 , CD49d claim 1 , CD58 claim 1 , CD61 claim 1 , CD71 claim 1 , CD142 claim 1 , CD146 claim 1 , CD201 claim 1 , CD340 claim 1 , and HLA-A/-B/-C.4. The cell of characterized in that the cell is measured negative for at least one marker selected from(a) CD26, CD49a, CD49d, CD58, CD61, CD71, CD142, CD146, CD201, CD340, and HLA-A/-B/-C; and/or(b) one or more of CD45, CD117, CD34, and HLA-DR.5. The cell of characterized in that the cell is further measured:(a) positive for at least one hepatic marker selected from albumin, HNF-4, and CYP3A4;(b) positive for at least one mesenchymal marker selected from Vimentin, a-smooth muscle actin (ASMA); and(c) negative for cytokeratin-19 (CK-19).6. The cell of characterized in that the cell is measured:(a) positive for CD13, CD73, CD90, CD105, CD29, CD44, CD47, CD49b, CD49C, CD49e, CD147, CD9, CD63, CD81, CD151, CD98, CD140b, β2-microglobulin, CD54, CD164, CD165, CD166, CD46, CD55, ...

ISOLATED LIVER STEM CELLS

A method of conducting in vitro toxicity testing is disclosed which includes the steps of exposing to a test agent a population of human liver progenitor or stem cells originated from human adult liver. The cells have the characteristics that the isolated human progenitor or stem cells express at least the mesenchymal markers vimentin and α-smooth muscle actin (ASMA), the hepatocyte marker albumin (ALB), are negative for cytokeratin-19 (CK-19), and have mesenchymal-like morphology. One or more effects, if any, of the test agent on the population of human liver progenitor or stem cells are observed. Also disclosed are methods of conducting in vitro drug metabolism studies by exposing to a test agent a population of the human liver progenitor or stem cells and methods of testing infected human liver progenitor or stem cells for effects of a test agent. 1. A method of conducting in vitro toxicity testing comprising: (i) the isolated human progenitor or stem cells express at least the mesenchymal markers vimentin and α-smooth muscle actin (ASMA),', '(ii) the isolated human progenitor or stem cells express the hepatocyte marker albumin (ALB),', '(iii) the isolated human progenitor or stem cells are negative for cytokeratin-19 (CK-19), and', '(iv) the isolated human progenitor or stem cells have mesenchymal-like morphology; and, '(a) exposing to a test agent a population of human liver progenitor or stem cells originated from human adult liver wherein the cells comprise the following characteristicsobserving at least one effect, if any, of the test agent on the population of human liver progenitor or stem cells.2. The method of claim 1 , wherein the at least one effect comprises an effect on cell viability claim 1 , cell function claim 1 , or both.3. The method of claim 1 , wherein the method further comprises infecting the population of human liver progenitor or stem cells originated from human adult liver with an infectious agent to provide an infected population of ...

Method for Measuring Bile Salt Export Transport and/or Formation Activity

A method is provided to measure modulation of bile salt export transport and/or formation activity in hepatocyte or stable cell line preparations by test agents including but not limited to drugs, drug candidates, biologicals, food components, herb or plant components, proteins, peptides, DNA, RNA. Furthermore, the method is to determine modulation of bile salt export transport and/or formation activity not only by said test agents, but further their metabolites or biotransformed products formed in situ. The bile salt export transport and/or formation activity modulation includes but not limited to inhibition, induction, activation and/or regulation. The method can be practiced to identify test agents, which have potential to cause liver injury, drug-drug interactions, and/or can be used as therapeutic agents for the treatment of cholestasis, abnormality of bile salt metabolism, liver diseases and cholesterol abnormality.

Immune hepatotoxicity screening method using hepatocytes derived from human stem cells

The present invention relates to an immune hepatotoxicity screening method using hepatocytes derived from human stem cells. After hepatocytes differentiated from human stem cells and human hepatocytes are treated with ethanol, CCl 4 , and acetaminophen to induce immune hepatotoxicity, a hepatocellular immunotoxic material assay system is constructed in order to verify cytokines, chemokines, and lipid mediators, which are mediators secreted from the hepatocytes, and an immunotoxic material can be confirmed in the cells having the induced hepatotoxicity by using the system. Therefore, the immune hepatotoxicity screening method using hepatocytes derived from human stem cells can be favorably used.

SCREENING METHOD FOR SELECTED AMINO-LIPID-CONTAINING COMPOSITIONS

The invention features a method of identifying therapeutically relevant compositions which include a therapeutic agent and 2,2-dimethylaminomethyl-[1-3]-dioxolane by screening for an effect of the agent on the liver of a model subject. 1. A method of evaluating a composition that includes a therapeutic agent and 2 ,2-Dilinoley 1-4-dimethylaminomethyl-[1 ,3]-dioxolane comprising:providing a composition that includes a therapeutic agent and 2,2-Dilinoley 1-4-dimethylaminomethyl-[1,3]-dioxolane;administering the composition to a test animal; anddetermining the effect of the composition on the expression of a target gene expressed in the liver of the animal,thereby evaluating the composition.2. The method of claim 1 , wherein the therapeutic agent is an RNA-based construct.3. The method of claim 2 , wherein the RNA-based construct is a dsRNA.4. The method of claim 1 , wherein the target gene is Factor VII.5. The method of claim 1 , wherein determining the effect of the composition comprises determining target protein levels.6. The method of claim 1 , wherein determining the effect of the composition comprises determining target mRNA levels.7. The method of claim 5 , wherein the level of target protein in blood is determined.8. The method of claim 6 , wherein the level of target mRNA in liver is determined.9. The method of claim 1 , further comprising comparing expression of the target gene with a preselected reference value.10. The method of claim 1 , wherein the composition further comprises a third component.11. The method of claim 1 , wherein the therapeutic agent is an antisense RNA claim 1 , ribozyme or microRNA.12. The method of claim 1 , wherein the test animal is a rodent.13. The method of claim 1 , wherein the test animal is a mouse.14. The method of claim 1 , wherein the composition reduces FVII protein or mRNA levels in the blood.15. The method of claim 1 , wherein the composition reduces FVII protein or mRNA levels in the liver.16. The method of claim 1 , ...

NOVEL RECOMBINANT HBV REPORTER SYSTEM

The present invention discloses a method for assessing the capacity of a substance to treat or prevent hepadnavirus infection. A reporter virus carrying genetic information for a first fragment of a recombinase and a reporter cell expressing a second fragment of the recombinase are used. When the reporter virus infects the reporter cell, the two fragments of the recombinase associate and excise a stop cassette that is flanked by two recombination sites and blocks the expression of a reporter gene. Accordingly, the present invention relates to a method of assessing the capacity of a substance to treat or prevent hepadnavirus infection, a hepadnavirus comprising a nucleic acid encoding a first fragment of a recombinase and a mammalian hepatocyte or hepatoma cell comprising a nucleic acid encoding a second fragment of a recombinase and a nucleic acid comprising a stop cassette flanked by two recombination sites fused to a reporter gene. 1. A method of assessing the capacity of a substance to treat or prevent hepadnavirus infection comprising:(a) Contacting a reporter cell with a reporter virus; and(b) Contacting the reporter cell with the substancewherein the reporter virus comprises a nucleic acid encoding a first fragment of a recombinase;wherein the reporter cell comprises a nucleic acid encoding a second fragment of the recombinase;wherein the first fragment of the recombinase and the second fragment of the recombinase are capable of forming a functional recombinase;wherein the reporter cell comprises a nucleic acid comprising a stop cassette flanked by two recombination sites fused to a reporter gene, wherein the presence of the stop cassette suppresses expression of the reporter gene.2. The method of claim 1 , wherein the hepadnavirus and/or the reporter virus is a hepatitis B virus.3. The method of claim 1 , wherein the reporter virus comprises a genome comprising a gene encoding an N-terminal fragment of Cre recombinase.4. The method of claim 1 , wherein the ...

METHOD AND SYSTEM FOR DETERMINING AT LEAST ONE PARAMETER OF INTEREST OF A MATERIAL

A method for determining at least one parameter of interest of a material comprises directing, using a radio frequency (RF) applicator, one or more RF energy pulses into a region of interest, the region of interest comprising a material having a parameter of interest and at least one reference, the material and the reference separated by at least one boundary; detecting, using an acoustic receiver, at least one multi-polar acoustic signal generated in the region of interest in response to the RF energy pulses; processing the at least one multi-polar acoustic signal to determine an electric field strength at the boundary; calculating a voltage standing wave ratio (VSWR) of the one or more RF energy pulses; and determining the at least one parameter of interest of the material based at least on the determined electric field strength and the VSWR. 1. A method for determining at least one parameter of interest of a material , the method comprising:directing, using a radio frequency (RF) applicator, one or more RF energy pulses into a region of interest, the region of interest comprising a material having a parameter of interest and at least one reference, the material and the reference separated by at least one boundary;detecting, using an acoustic receiver, at least one multi-polar acoustic signal generated in the region of interest in response to the RF energy pulses;processing the at least one multi-polar acoustic signal to determine an electric field strength at the boundary;calculating a voltage standing wave ratio (VSWR) of the one or more RF energy pulses; anddetermining the at least one parameter of interest of the material based at least on the determined electric field strength and the VSWR.2. The method of wherein the at least one parameter of interest is one of a Gruneisen Parameter of the material and a conductivity of the material.3. The method of claim 2 , wherein the at least one parameter of interest is a product of the Gruneisen Parameter of the ...

MULTI-WELL MICROPATTERNING BY ABLATION

The present invention is drawn to the generation of micropatterns of biomolecules and cells on standard laboratory materials through selective ablation of a physisorbed biomolecule with oxygen plasma. In certain embodiments, oxygen plasma is able to ablate selectively physisorbed layers of biomolecules (e.g., type-I collagen, fibronectin, laminin, and Matrigel) along complex non-linear paths which are difficult or impossible to pattern using alternative methods. In addition, certain embodiments of the present invention relate to the micropatterning of multiple cell types on curved surfaces, multiwell plates, and flat bottom flasks. The invention also features kits for use with the subject methods. 1. A method of forming a micropatterned substrate , comprising the steps of:adsorbing molecules onto a surface of a substrate, thereby forming a coated surface of the substrate;comprising a micropatterned etch mask onto the coated surface of said substrate; andexposing the compressed micropatterned etch mask and coated surface of the substrate to a gas plasma for a period of time, thereby ablating the exposed surfaces of the substrate.2. The method of claim 1 , further comprising rinsing and drying said coated surface after the adsorbing step.3. The method of claim 1 , wherein said exposing step is carried out in a plasma asher.4. The method of claim 1 , wherein the micropatterned etch mask is one solid elastomericpiece.5. The method of claim 1 , wherein the micropatterned etch mask comprises a plurality of pillars.6. The method of claim 1 , wherein the micropatterned etch mask comprises chrome or elastomeric poly(dimethylsiloxane) or rubber or plastic.7. The method of claim 1 , wherein the adsorbed molecules are different.8. The method of claim 7 , wherein the different molecules each have a different pattern.9. The method of claim 1 , wherein the micropatterned etch mask comprises plastic.10. The method of claim 1 , wherein the micropatterned etch mask comprises an about ...

USES AND METHODS FOR MODULATING BILE ACID HOMEOSTASIS AND TREATMENT OF BILE ACID DISORDERS AND DISEASES

The invention relates to variants and fusions of fibroblast growth factor 19 (FGF19), variants and fusions of fibroblast growth factor 21 (FGF21), fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21), and variants or fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21) proteins and peptide sequences (and peptidomimetics), having one or more activities, such as bile acid homeostasis modulating activity, and methods for and uses in treatment of bile acid and other disorders. 171-. (canceled)72. A method of reducing CYP7a1 expression in a subject having nonalcoholic steatohepatitis (NASH) , comprising administering to the subject an effective amount of a peptide , wherein the peptide comprises:a) an N-terminal region comprising at least seven amino acid residues, the N-terminal region having a first amino acid position and a last amino acid position, wherein the N-terminal region comprises DSSPL (SEQ ID NO:121) or DASPH (SEQ ID NO:122); and (i) a first C-terminal region sequence comprising WGDPIRLRHLYTSG (amino acids 16 to 29 of SEQ ID NO:99 [FGF19]), wherein the W residue corresponds to the first amino acid position of the C-terminal region; and', '(ii) a second C-terminal region sequence comprising PHGLSSCFLRIRADGVVDCARGQSAHSLLEIKAVALRTVAIKGVHS VRYLCMGADGKMQGLLQYSEEDCAFEEEIRPD GYNVYRSEKHRL PVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESD MFSSPLETDSMDPFGLVTGLEAVRSPSFEK (amino acid residues 30 to 194 of SEQ ID NO:99 [FGF19]); or a sequence comprising from 1 to 5 amino acid substitutions, deletions or insertions thereof;, 'b) a C-terminal region comprising a portion of SEQ ID NO:99 [FGF19], the C-terminal region having a first amino acid position and a last amino acid position, wherein the C-terminal region comprises'}wherein the peptide(A) has reduced hepatocellular carcinoma (HCC) formation as compared to FGF19, or as compared to an FGF19 variant sequence having any of GQV, GDI, WGPI (SEQ ID NO:171 ...

ASSAY-READY RECOMBINANT CELLS TRANSIENTLY OVEREXPRESSING GENES ENCODING DRUG TRANSPORTER PROTEINS AND/OR DRUG METABOLIZING ENZYMES

Recombinant cells including one or more transiently overexpressed genes encoding a drug transporter protein, wherein the recombinant cell is cryopreserved and activity of the drug transporter protein is detectable in a population of the recombinant cells prior to cryopreservation at an uptake ratio of at least 5. Processes of preparing cryopreserved transiently transfected recombinant cells, including transiently transfecting cells with one or more genes encoding a drug transporter protein and cryopreserving the transiently transfected recombinant cells within 48 hours of transfection. A population of the transiently transfected recombinant cells transiently overexpress the one or more genes encoding the drug transporter protein at a detectable level prior to cryopreserving and the detectable level is an uptake ratio of at least 5. 1. A recombinant cell comprising one or more transiently overexpressed genes encoding a drug transporter protein , wherein:the recombinant cell is cryopreserved, andactivity of the drug transporter protein is detectable in a population of the recombinant cells prior to cyropreservation at an uptake ratio of at least 5.2. The recombinant cell of claim 1 , wherein the activity of the drug transporter protein would be detectable in a population of the recombinant cells following thaw from cryopreservation at an uptake ratio of at least 5.3. The recombinant cell of claim 2 , wherein the activity of the drug transporter protein would be detectable in the population of the recombinant cells following thaw from cryopreservation at an uptake ratio of from about 5 to about 150.4. The recombinant cell of claim 1 , wherein:the activity of the drug transporter protein would be detectable in a plated population of the recombinant cells following thaw from cryopreservation at an uptake ratio of from about 5 to about 30 within 4 hours of thawing.5. The recombinant cell of claim 1 , wherein:the activity of the drug transporter protein would be detectable ...

INTERMITTENT STARVATION TECHNIQUES TO PROLONG FUNCTIONAL LIFETIME OF LIVER CELLS IN CULTURE

The present disclosure relates to compositions and methods for culturing a population of hepatocytes in vitro, comprising co-culturing the population of hepatocytes with at least one non-parenchymal cell population and incubating the co-culture in culture medium, wherein the co-culture is periodically incubated in culture medium that does not comprise serum (serum-free culture medium). 1. A method of culturing a population of hepatocytes in vitro , comprising co-culturing the population of hepatocytes with at least one non-parenchymal cell population and incubating the co-culture in culture medium ,wherein the cell populations are disposed in a micropattern on a culture substrate and the micropattern comprises a predetermined two-dimensional pattern of multiple microdots, the micropattern defined by a microdot diameter and a center-to-center spacing between each of any two neighboring microdots, and the microdots comprise the hepatocytes and the space between the microdots comprises the non-parenchymal cell population, andwherein the co-culture is incubated in culture medium comprising serum for at least one time period of about 1 hour to about 6 weeks and is incubated in culture medium that does not comprise serum (serum-free culture medium) for at least one time period of about 1 hour to about 6 weeks.2. The method according to claim 1 , wherein each microdot has a diameter of about 500 μm and the center-to-center spacing between each of any two neighboring microdots is about 1200 μm.3. The method according to claim 1 , wherein the composition is incubated in culture medium comprising serum for at least one time period of at least 1 week before the composition is incubated in serum-free culture medium.4. The method according to claim 1 , wherein the co-culture is incubated in culture medium comprising serum for at least one time period of at least 2 weeks before the composition is incubated in serum-free culture medium.5. The method according to claim 1 , wherein ...

METHOD FOR HIGH-THROUGHPUT SCREENING OF COMPOUNDS AND COMBINATIONS OF COMPOUNDS FOR DISCOVERY AND QUANTIFICATION OF ACTIONS, PARTICULARLY UNANTICIPATED THERAPEUTIC OR TOXIC ACTIONS, IN BIOLOGICAL SYSTEMS

The invention enables high-throughput screening of compounds in living systems to detect unanticipated or unintended biological actions. The invention also allows for screening, detection, and confirmation of new indications for approved drugs. Screening and detection of toxic effects of compounds also can be achieved by using the methods of the invention. The methods comprise administering isotope-labeled substrates to a living system so that the label is incorporated into molecules in a manner that reveals flux rates through metabolic pathways thought to be involved in a disease. Comparisons between living systems exposed to compounds and living systems not so exposed reveals the effects of the compounds on the flux rates through the metabolic pathways. Combinations or mixtures of compounds can be systematically screened to detect unantidpated or unintended biological actions, including synergistic actions, in the same manner. 114-. (canceled)15. A method for high-throughput screening (HTS) of combinations or mixtures of two or more compounds for actions on molecular flux rates in liver collagen synthesis and breakdown , said method comprising:a) administering said two or more compounds to a living system;b) administering an isotope-labeled substrate to said living system for a period of time sufficient for said isotope-labeled substrate to enter into and pass through liver collagen synthesis and breakdown and thereby enter into and label a targeted molecule or molecules of interest within said liver collagen synthesis and breakdown in said living system, wherein the isotope-labeled substrate is stable isotope-labeled water and the targeted molecule or molecules of interest is selected from the group consisting of proteins, peptides, and amino acids;c) obtaining one or more samples from said living system, wherein said one or more samples comprise one or more isotope-labeled targeted molecules of interest;d) measuring the isotopic content or the isotopic pattern ...

Treatment

The present invention derives from the unexpected finding that ADRA2a antagonists decrease fibrosis in non-alcoholic fatty liver disease (NAFLD). Thus, by antagonising ADRA2a, many of the unwanted consequences or symptoms of NAFLD, may be reduced, such as impaired cognitive activity and fibrosis progression. The present invention utilises these findings to identify and provide ADRA2a antagonists that may be used in the treatment of fibrosis in NAFLD.

DIRECT CONVERSION METHOD OF SOMATIC CELL INTO HEPATIC STEM CELL, HEPATIC CELL, OR CHOLANGIOCYTE

The present invention relates to a composition for inducing direct conversion from a somatic cell into one or more kinds selected from the group consisting of an induced Hepatic stem cell (iHSC), a hepatocyte, and a cholangiocyte, and a method of direct conversion of a somatic cell into one or more kinds selected from the group consisting of an induced Hepatic stem cell, a hepatocyte, and a cholangiocyte. 1. A method of direct conversion from a somatic cell to at least one selected from the group consisting of a hepatic stem cell , a hepatocyte , and a cholangiocyte , the method comprising a step of introducing a composition for inducing direct conversion into the somatic cell ,wherein the composition comprises at least one selected from the group consisting of:(1) OCT4 (Octamer-binding transcription factor 4) protein, HNF4α (Hepatocyte nuclear factor 4-alpha) protein, NR4A2 (Nuclear receptor subfamily 4 group A member 2) protein, NR4A1 (Nuclear receptor subfamily 4 group A member 1) protein, TBX3 (T-box transcription factor 3) protein, NR5A1 (Nuclear receptor subfamily 5 group A member 1) protein, NR5A2 (Nuclear receptor subfamily 5 group A member 2) protein, and NR0B2 (Nuclear receptor subfamily 0 group B member 1) protein,(2) nucleic acid molecules encoding each of the proteins, and(3) vectors into which each of the nucleic acid molecules is introduced.2. The method of claim 1 , wherein the composition for inducing direct conversion comprises:at least one selected from the group consisting of OCT4 protein, a nucleic acid molecule encoding OCT4 protein, and a vector into which the nucleic acid molecule encoding OCT4 protein is introduced, andat least one selected from the group consisting of HNF4α protein, NR4A1 protein, NR4A2 protein, a nucleic acid molecule encoding HNF4α protein, a nucleic acid molecule encoding NR4A1 protein, a nucleic acid molecule encoding NR4A2 protein, a vector into which the nucleic acid molecule encoding HNF4α protein is introduced, a ...

Method for rapid identification of pharmacologically active chemical entities associated with the efficacy of ethnobotanical substances

The method includes the steps of performing in-vitro liver, intestinal and/or expressed enzyme assays with selected ethnobotanical substances, for both humans and a variety of animal species, to produce an array of resulting chemical entities, such as metabolites, for the human and the animals. Comparisons are then made between the chemical entities from the human in-vitro studies and the animal in-vitro studies to determine the closest match. The animal with the closest match is then used for an in-vivo study. If a match is present between the animal in-vivo results and the human in-vitro results, the matched chemical entity is isolated or synthesized and then further tested to determine the suitability of the matched chemical entity as a treatment drug.

In vitro drug metabolism reagent and uses thereof

The present disclosure provides an in vitro reagent for evaluating xenobiotic metabolism in a cell culture based assay. The in vitro reagent is an admixture of metabolically competent cells and exogenous drug metabolizing enzyme co-factors follow by cryopreservation in the absence of cryopreservation agent so that the cells would be rendered permeable upon thawing due to plasma membrane disruption (while maintaining the integrity of organelles). The permeabilized plasma membranes allow ready diffusion of the exogenous cofactors into the cells to enhance the activities of cellular drug metabolizing enzymes. Addition of a xenobiotic test compound to the thawed in vitro reagent allows metabolism of the test compound by the metabolically competent cells, with metabolites readily diffusible outside the cells due to the permeabilized plasma membranes.

Componential Analyzer, Drug Efficacy Analyzer, and Analysis Method

Application of the present invention enables quantification of fractions of candidate pharmaceutical compounds (a parent compound and its metabolites), one excreted to the basolateral (Basal/Basolateral)-side via transporters and by diffusion, one excreted to the lumen (Apical)-side, and one remained in the cells. This enables determination of the total amount of the administered candidate pharmaceutical compounds and the distribution ratio of the fractions. The kinetics of the administered candidate pharmaceutical compounds can be evaluated, thereby enabling in vitro screening of an enormous number of candidate pharmaceutical compounds for drug candidates exhibiting the efficacy. The object of the present invention is to provide an apparatus and method for understanding a total picture of pharmacokinetics in vitro by quantifying a fraction of basolateral (Basal/Basolateral) efflux, a fraction of lumen (Apical)-side excretion, and a fraction remaining in a cell of a drug which has been administered to the cell to determine the distribution ratio of each fraction. 1. A method of componential analysis , comprising:a temperature controlling step of controlling a first temperature in a first container and a third container containing a cell and a second temperature in a second container containing the cell so that the second temperature is lower than the first temperature;a measuring step of measuring:an amount of a component excreted from the cell in the first container to the first buffer solution in the first container,an amount of the component excreted from the cell in the second container to the first buffer solution in the second container, andan amount of the component excreted from the cell in the third container to a second buffer solution comprising a reagent for removing cell-cell adhesion of the cell in the third container; andan analyzing step of analyzing, based on the result measured in the measuring step, each of:an amount of the component excreted via ...

PRODUCTION OF VIRUS-RECEPTIVE PLURIPOTENT STEM CELL (PSC)-DERIVED HEPATOCYTES

The present disclosure provides methods for maturing hepatocytes comprising culturing with cyclic adenosine monophosphate and a Janus kinase inhibitor. There is also provided a method for screening inhibitors of hepatitis B virus infection and/or replication. 1. An in vitro method of producing virus-receptive pluripotent stem cell-derived hepatocytes comprising:(a) obtaining pluripotent stem cell (PSC)-derived hepatocytes; and(b) culturing the PSC-derived hepatocytes in media comprising cyclic adenosine monophosphate (cAMP) and a Janus kinase inhibitor (JAKi), thereby producing virus receptive hepatocytes.2. The method of claim 1 , wherein the cAMP and JAKi are cultured sequentially in different media.3. The method of claim 2 , wherein the cAMP is administered in a first medium and the JAKi is administered in a second medium.4. The method of claim 1 , wherein the pluripotent stem cell is human.5. The method of claim 1 , wherein the pluripotent stem cell is an embryonic stem cell.6. The method of claim 1 , wherein the pluripotent stem cell is an induced pluripotent stem cell.7. The method of claim 1 , wherein the JAKi is 2-(1 claim 1 ,1-Dimethylethyl)-9-fluoro-3 claim 1 ,6-dihydro-7H-benz[h]-imidaz[4 claim 1 ,5-f]isoquinolin-7-one (JAK Inhibitor 1) claim 1 , 1 claim 1 ,2 claim 1 ,3 claim 1 ,4 claim 1 ,5 claim 1 ,6-Hexabromocyclohexane claim 1 , or (E)-4 claim 1 ,4′-(1 claim 1 ,2-Diethyl-1 claim 1 ,2-ethenediyl)bis[2-[(diethylamino)methyl]-phenol (9CI) claim 1 , 4 claim 1 ,4′-(3E)-Hex-3-ene-3 claim 1 ,4-diylbis{2-[(diethylamino)methyl]phenol} (NSC33994).8. The method of claim 1 , wherein the JAKi is present at a concentration of about 0.1 μM to about 5 μM.9. The method of claim 8 , wherein the JAKi is present at a concentration of about 1 μM.10. The method of claim 1 , wherein the cAMP is present at a concentration of about 0.1 mM to about 3 mM.11. The method of claim 10 , wherein the cAMP is present at a concentration of about 1 mM.12. The method of claim 1 , wherein ...

HUMAN FATTY-LIVER MODEL CELLS

An object of the present invention is to provide human fatty-liver model cells showing symptoms of the hepatic tissue of fatty liver. The present invention relates to human fatty-liver model cells, which are produced by culturing human hepatocytes derived from fatty liver in a medium containing dimethyl sulfoxide. 1. A method for producing human fatty-liver model cells , comprising a step of culturing human hepatocytes derived from fatty liver in a medium containing dimethyl sulfoxide.2. The method according to claim 1 , wherein the human hepatocytes derived from fatty liver are collected from a chimeric non-human animal having human hepatocytes.3. The method according to or claim 1 , wherein the culturing is carried out for more than 3 days.4. Human fatty-liver model cells that secrete and/or accumulate lipid.5. The cells according to claim 4 , comprising a lipoprotein including a very low density lipoprotein (VLDL) and a low density lipoprotein (LDL) claim 4 , wherein VLDL is comprised more than LDL.6. The cells according to claim 4 , having increased expression of a fatty liver related gene.7. The cells according to claim 6 , wherein the fatty liver related gene is at least one gene selected from the group consisting of FASN claim 6 , SREBF1 and G6PC.8. A method for screening for a substance effective for human fatty liver claim 6 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'administering a test substance to the cells according to ; and'}comparing severity of fatty-liver symptoms between cells to which the test substance is administered and cells to which the test substance is not administered.9. A method for evaluating toxicity of a test substance to human fatty liver claim 6 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'administering a test substance to the cells according to ; and'}comparing survival rate and severity of fatty-liver symptoms between cells to which the test substance is administered ...

METHODS FOR CHARACTERIZING TIME-BASED HEPATOTOXICITY

Methods of characterizing the time-based hepatotoxicity of a test compound are provided. In some embodiments the provided methods allow prediction of the hepatotoxic potential of a test compound in a way that is improved relative to that achievable with prior art methods. In some embodiments the methods may be used to quantify the relationship between measurements or estimates of toxicity made at different, successive points in time, to provide highly meaningful tool to assess the likely in vivo hepatotoxicity of test compounds in vivo. 1. A method of characterizing the time-based hepatotoxicity of a test compound , comprising:a) incubating a first in vitro culture comprising hepatocytes with a test compound for a first culture period;b) measuring at least one cytotoxic effect of the test compound on the hepatocytes of the first in vitro culture over the first culture period to thereby define the hepatotoxicity of the test compound over the first culture period;c) incubating a second in vitro culture comprising hepatocytes with the test compound for a second culture period that is longer than the first culture period;d) measuring at least one cytotoxic effect of the test compound on the hepatocytes of the second in vitro culture over the second culture period to thereby define the hepatotoxicity of the test compound over the second culture period; ande) comparing the hepatotoxicity of the test compound over the first culture period to the hepatotoxicity of the test compound over the second culture period to thereby characterize the time-based hepatotoxicity of the test compound.2. The method of claim 1 , wherein the hepatotoxicity of the test compound over the second culture period is greater than the hepatotoxicity of the test compound over the first culture period and the test compound is identified as exhibiting time-based hepatotoxicity.3. The method of claim 1 , wherein the hepatotoxicity of the test compound over the second culture period is greater than the ...

Hepatocyte induction method

A method for producing hepatocytes from hepatoblasts is provided. The method includes the step of culturing the hepatoblasts in a medium containing a compound selected from the group consisting of pregnenolone and an adrenergic agonist. The hepatoblasts can be obtained by culturing endodermal cells in a medium containing DMSO, and the endodermal cells can be obtained by culturing pluripotent stem cells in a medium containing Activin A and a GSK-3β inhibitor. Accordingly, a method for producing hepatocytes from pluripotent stem cells is also provided by employing the method of the present invention.

CLINICAL GENE SIGNATURE-BASED HUMAN CELL CULTURE MODEL AND USES THEREOF

The present invention provides a simple and robust human liver cell-based system in which persistent hepatitis C infection, persistent hepatitis B infection or ethanol exposure induces a clinical Prognostic Liver Signature (PLS) high-risk gene signature. The cellular model system for hepatocellular carcinoma (HCC)/cirrhosis development and progression may be used in the screening of compounds useful in the treatment and/or prevention of cirrhosis and/or HCC as well as in the identification biomarkers for the prediction of liver disease (especially cirrhosis) progression and HCC. The present invention also relates to specific compounds that have been identified, using such screening methods, as useful in the treatment and/or the prevention of HCC/cirrhosis. 1. A method for generating a cellular model for cirrhosis and/or hepatocellular carcinoma (HCC) development and progression , said method comprising steps of:(a) differentiating liver cancer cells to obtain hepatocyte-like cells; and(b) submitting said hepatocyte-like cells to one hepatocarcinogenic agent to obtain liver cells exhibiting a Prognostic Liver Signature (PLS) high-risk gene signature.2. The method according to claim 1 , wherein the PLS high-risk gene signature is the PLS high-risk 186-gene signature presented in Table la claim 1 , wherein the 73 high-risk genes claim 1 , or a subset thereof claim 1 , are overexpressed and the 113 low-risk genes claim 1 , or a subset thereof claim 1 , are underexpressed.3. The method according to claim 1 , wherein said liver cancer cells are primary cells isolated from a liver cancer tissue sample or cells from a liver cancer cell line.4. The method according to claim 3 , wherein the liver cancer cell line is selected from the group consisting of the Huh7.5.1 claim 3 , Hep3B.1-7 claim 3 , HepG2 claim 3 , HepG2-NTCP claim 3 , HepG2AD38 claim 3 , HepG2215 claim 3 , SkHepI claim 3 , C3A claim 3 , PLC/PRF/5 and SNU-398 cell lines.5. The method according to claim 1 , ...

UROKINASE-TYPE PLASMINOGEN ACTIVATOR TRANSGENIC MOUSE

The present invention provides a mouse with liver damage, having a high degree of damage against the mouse's original hepatocytes while having a uPA gene in a heterozygous form, and a method for efficiently preparing the mouse. Specifically, the method for preparing a mouse with liver damage having the uPA gene in a heterozygous form comprises the following steps of: 125-. (canceled)26. A method for preparing a mouse with liver damage , which has a urokinase-type plasminogen activator (uPA) gene in a heterozygous form , comprising the following steps of:(i) transforming mouse ES cells with a DNA fragment containing a liver-specific promoter/enhancer and cDNA that encodes uPA operably linked under the control thereof;(ii) injecting the transformed mouse ES cells obtained in step (i) into a host embryo;(iii) transplanting the host embryo obtained in step (ii) via the injection of the ES cells into the uterus of a surrogate mother mouse, so as to obtain a chimeric mouse;(iv) crossing the chimeric mice obtained in step (iii), so as to obtain a transgenic mouse in which the DNA fragment is introduced in a heterozygous form; and(v) obtaining a transgenic mouse, wherein the serum ALT level increases at least from when it is 6 weeks old to when it is 8 weeks old.27. A method for preparing a mouse with liver damage that has a urokinase-type plasminogen activator (uPA) gene in a heterozygous form , comprising the following steps of:(i) transforming mouse ES cells with a DNA fragment containing a liver-specific promoter/enhancer and cDNA that encodes uPA operably linked under the control thereof;(ii) injecting the transformed mouse ES cells obtained in step (i) into a host embryo;(iii) transplanting the host embryo obtained in step (ii) via the injection of the ES cells into the uterus of a surrogate mother mouse, so as to obtain a chimeric mouse;(iv) crossing the chimeric mice obtained in step (iii), so as to obtain a transgenic mouse into which the DNA fragment is introduced ...

Microfluidic platform for target and biomarker discovery for non-alcoholic fatty liver disease

A method for developing stratified medicine for nonalcoholic fatty liver disease (NAFLD includes obtaining a microphysiological system (MPS) comprising a liver tissue cytoarchitecture, adipose tissue, or both. The method includes inducing metabolic dysfunction representing NAFLD in the liver or adipose tissue of the MPS. The method includes generating, based on inducing the metabolic dysfunction, transcriptomics data for the MPS. The method includes applying a drug to the MPS using a dosing regimen. The method includes monitoring changes in the transcriptomics data based on applying the drug. The method includes generating a model relating the changes in the transcriptomics data to the dosing regimen of the drug.

METHOD FOR PREVENTING OBESITY-INDUCED FATTY LIVER BY INHIBITING KCTD17

The present invention provides methods for reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that decreases KCTD17 expression in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels. 1. A method of reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that decreases KCTD17 expression in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels.2. A method of reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that prevents PHLPP2 degradation in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels.3. The method of claim 2 , wherein the pharmaceutical composition inhibits Glucagon signaling.4. A method of reducing a subject's hepatic and plasma triglyceride levels comprising administering to a subject in need thereof a pharmaceutical composition comprising a pharmaceutical carrier and a compound that inhibits Glucagon signaling in liver cells in an amount effective to reduce the subject's hepatic and plasma triglyceride levels.5. The method of claim 4 , wherein the pharmaceutical composition reduces PHLPP2 degradation.6. The method of or claim 4 , wherein the pharmaceutical composition increases free Raptor in the liver cells.7. The method of - claim 4 , wherein the pharmaceutical composition decreases PHLPP2 phosphorylation at Serine 1119 and Serine 1210 residues in liver cells.8. The method of claim 6 , wherein the pharmaceutical composition prevents PHLPP2 degradation in liver cells.9. The method of any one of - claim 6 ...

IN VITRO MODEL OF MACROSTEATOTIC (FATTY) LIVER

The present invention relates to a system and methods for identifying a compound for de-fatting and functional recovery of macrosteatotic hepatocytes. 1. A culture system , comprising a cell population comprising cultured macrosteatotic hepatocytes from an animal , and a culture medium.2. The culture system of claim 1 , wherein the cell population further comprises cells selected from the group consisting of fibroblasts claim 1 , Kupffer cells claim 1 , and cells from a non-liver organ or tissue.3. The culture system of claim 2 , wherein the macrosteatotic hepatocytes are maintained in a collagen matrix.4. The culture system of claim 3 , wherein the collagen matrix is in a sandwich configuration.5. The culture system of claim 1 , wherein the animal is a non-human mammal.6. The culture system of claim 5 , wherein the non-human mammal is a rodent.7. The culture system of claim 1 , wherein the animal is a human.8. The culture system of claim 1 , wherein the macrosteatotic hepatocytes contain lipid macrodroplets.9. The culture system of claim 1 , wherein the culture medium is selected from the group consisting of a standard medium claim 1 , a steatosis inducing medium claim 1 , and a steatosis reducing medium.10. A screening method of identifying a compound or composition for de-fatting and functional recovery of macrosteatotic hepatocytes claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'obtaining a first culture system of , having a population of cultured macrosteatotic hepatocytes;'}incubating the macrosteatotic hepatocytes in a test medium containing a test compound or test composition for a first period of time; anddetermining a macrosteatosis level or a function level of the macrosteatotic hepatocytes,wherein the test compound is effective for de-fatting and functional recovery of macrosteatotic hepatocytes if (i) the macrosteatosis level is lower than a control macrosteatosis level or (ii) the function level is higher than a control function ...

MASS SPECTROMETRY QUANTITATION OF P450 ISOFORMS IN HEPATOCYTES

A method for screening a drug for cytochrome P450 (CYP) induction is provided and can include incubating the drug with a microsome-containing biological sample and then quantitating at least one cytochrome P450 isoform. The isoforms can be selected from 2B6, 3A4, 1A2, and 3A5 isoforms. In some embodiments, the method uses liquid chromatography tandem mass spectrometry (LC-MSMS). A quantitated value can be compared to a threshold value and the drug can be determined to exhibit an acceptable CYP induction potential when the quantitated value does not exceed the threshold value. Isolated peptides are also provided. 1. A method for determining an amount of at least one isoform of cytochrome P450 (CYP) in a sample , said method comprising subjecting said sample to an analysis with a liquid chromatograph tandem mass spectrometer system , said system comprising a triple quadrupole instrument and Multiple Reaction Monitoring (MRM) , said analysis comprising detecting at least one isolated peptide of CYP 3A4 , CYP 1A2 or CYP 3A5.2. The method of wherein the isoform is CYP 3A4 and said isolated peptide has the amino acid sequence of SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 claim 1 , or SEQ ID NO: 73. The method of claim 2 , wherein the amino acid sequence is SEQ ID NO:5 and said analysis is performed by monitoring a precursor-product ion pair transition having an m/z value of about 440/549 claim 2 , 440/650 claim 2 , or 440/532 in said system and processing data from said monitoring to determine said amount.4. The method of claim 2 , wherein the amino acid sequence is SEQ ID NO:6 and said analysis is performed by monitoring a precursor-product ion pair transition having an m/z value of about 704/794 claim 2 , 704/929 claim 2 , 564/689 claim 2 , 564/745 claim 2 , or 564/790 in said system and processing data from said monitoring to determine said amount.5. The method of claim 2 , wherein the amino acid sequence is SEQ ID NO:7 and said analysis is performed by monitoring a precursor ...

METHOD FOR PRODUCING ADULT LIVER PROGENITOR CELLS

Novel adult liver progenitor cells (called H2Stem Cells) have been have been characterized on the basis of a series of biological activities and markers. Methods for producing H2Stem Cells allow providing such cells in the form of adherent cells and three-dimensional cell clusters in suspension that can be differentiated into cells having strong liver-specific activities and/or that can be used for treating liver diseases or for evaluating the efficacy, the metabolism, and/or toxicity of a compound. 131-. (canceled)32. A method for evaluating the efficacy , the metabolism , the stability , and/or the toxicity of one or more exogenous components , said method comprising: [ (i) at least one hepatic marker selected from albumin, HNF-3B, HNF-4, CYP1 A2, CYP2C9, CYP2E1 and CYP3A4;', '(ii) at least one mesenchymal marker selected from Vimentin, CD90, CD73, CD44, and CD29;', '(iii) at least one liver-specific activity selected from urea secretion, bilirubin conjugation, alpha-1 -antitrypsin secretion, and CYP3A4 activity;', '(iv) Sushi domain containing protein 2 (SUSD2); and', '(v) Cytokeratin-19 (CK-19);, '(I) a cell, wherein the cell is an adult liver progenitor cell characterized in that said cell is measured positive for, '(II) an isolated cell population comprising at least 60% or between 60% and 99% or between 70% or 90% of cells of part (I);', '(III) a biological material isolated from a cell of part (I) or population of part (II), wherein the biological material is a conditioned cell culture media, a protein extract, a membrane vesicle, or any fraction thereof comprising one or more isolated proteins, nucleic acids, metabolites, and/or antigens; or', '(IV) a composition comprising a cell of part (I), a cell population of part (II), or a biological material of part (III);, '(a) providing(b) exposing said cell, said cell population, said composition, or said biological material to one or more exogenous components selected from chemical compounds, proteins, nucleic ...

IN VITRO MODEL FOR PATHOLOGICAL OR PHYSIOLOGIC CONDITIONS

The present invention generally relates to in vitro methods for mimicking in vivo pathological or physiologic conditions. The methods comprise applying shear forces to a cell type or cell type plated on a surface within a cell culture container. Methods for testing drugs or compounds in such systems are also described. 1299-. (canceled)300. A method of mimicking a pathological or physiologic condition in vitro , the method comprising:adding a culture medium and at least one factor to a cell culture container;plating at least one cell type on at least one surface within the cell culture container, wherein the surface comprises a surface of a porous membrane suspended in the cell culture container; andapplying a shear force upon a second surface of the porous membrane, the shear force resulting from flow of the culture medium induced by a flow device, the flow mimicking flow to which the at least one cell type is exposed in vivo in the pathological or physiologic condition, wherein: (i) within the in vivo concentration range of the factor observed in the pathological condition; or', '(ii) within the concentration range of the factor that would result in vivo from administration of a drug or a compound; or, 'the concentration of the factor in the culture media for mimicking the pathological condition is either (i) within the in vivo concentration range of the factor observed in the physiologic condition; or', '(ii) within the concentration range of the factor that would result in vivo from administration of a drug or a compound., 'the concentration of the factor in the culture media for mimicking the physiologic condition is either301. The method of claim 300 , wherein the method further comprises testing a drug or a compound for an effect on the pathological or physiologic condition claim 300 , wherein testing the drug or the compound for the effect on the pathological or physiologic condition comprises:adding the drug or the compound to the culture medium after ...

IN VITRO METHOD FOR SCREENING TESTING COMPOUND TO EVALUATE ITS POTENTIAL AS LIVER DRUG

An in vitro method for screening a testing compound to evaluate its potential as a liver drug of the disclosure is provided. The method includes applying the testing compound to cells of an isolated human liver tumor cell line, named as ITRI-H28, measuring a cell viability of the cells and determining the effect of the testing compound on the cells by calculating a half maximal inhibitory concentration (IC) of the testing compound. 1. An in vitro method for screening a testing compound to evaluate its potential as a liver drug , comprising:(a) applying the testing compound to cells of an isolated human liver tumor cell line, named as ITRI-H28;(b) measuring a cell viability of the cells; and{'sub': '50', '(c) determining the effect of the testing compound on the cells by calculating a half maximal inhibitory concentration (IC) of the testing compound.'}2. The in vitro method as recited in further comprising comparing the effect of the testing compound on the cells to a control.3. The in vitro method as recited in claim 1 , wherein Sorafenib is used as the control.4. The in vitro method as recited in claim 1 , wherein the liver drug comprises a hepatitis C virus-related hepatocellular carcinoma (HCV-related HCC) drug.5. The in vitro method as recited in claim 1 , wherein the liver drug comprises a drug for liver failure.6. The in vitro method as recited in claim 1 , wherein the liver drug comprises a drug for liver cancer stem cell.7. The in vitro method as recited in claim 1 , wherein the isolated human liver tumor cell line further comprises a reporter gene.8. The in vitro method as recited in claim 7 , wherein the reporter gene expresses fluorescence or luminescence.9. The in vitro method as recited in claim 1 , wherein the isolated human liver tumor cell line secretes alpha-fetoprotein and albumin.10. The in vitro method as recited in claim 1 , wherein the isolated human liver tumor cell line has a cancer stem cell potential.11. The in vitro method as recited in ...

IN VITRO METHOD FOR SCREENING TESTING COMPOUND TO EVALUATE ITS POTENTIAL AS LIVER DRUG

An in vitro method for screening a testing compound to evaluate its potential as a liver drug is provided. The method includes applying the testing compound to cells of an isolated human liver tumor cell line, named as ITRI-H16, measuring a cell viability of the cells and determining the effect of the testing compound on the cells by calculating a half maximal inhibitory concentration (IC) of the testing compound. 1. An in vitro method for screening a testing compound to evaluate its potential as a liver drug , comprising:(a) applying the testing compound to cells of an isolated human liver tumor cell line, named as ITRI-H16;(b) measuring a cell viability of the cells; and{'sub': '50', '(c) determining the effect of the testing compound on the cells by calculating a half maximal inhibitory concentration (IC) of the testing compound.'}2. The in vitro method as recited in further comprising comparing the effect of the testing compound on the cells to a control.3. The in vitro method as recited in claim 1 , wherein Sorafenib is used as the control.4. The in vitro method as recited in claim 1 , wherein the liver drug comprises a hepatitis C virus-related hepatocellular carcinoma (HCV-related HCC) drug.5. The in vitro method as recited in claim 1 , wherein the liver drug comprises a drug for liver failure.6. The in vitro method as recited in claim 1 , wherein the liver drug comprises a drug for liver cancer stem cell.7. The in vitro method as recited in claim 1 , wherein the isolated human liver tumor cell line further comprises a reporter gene.8. The in vitro method as recited in claim 7 , wherein the reporter gene expresses fluorescence or luminescence.9. The in vitro method as recited in claim 1 , wherein the isolated human liver tumor cell line secretes alpha-fetoprotein and albumin.10. The in vitro method as recited in claim 1 , wherein the isolated human liver tumor cell line has a cancer stem cell potential.11. The in vitro method as recited in claim 1 , wherein ...

HEPATOCYTES AND HEPATIC NON-PARENCHYMAL CELLS, AND METHODS FOR PREPARATION THEREOF

The present invention pertains to hepatocytes, liver progenitor cells, cholangiocytes, liver sinusoidal endothelial progenitor cells, liver sinusoidal endothelial cells, hepatic stellate progenitor cells, hepatic stellate cells, and liver cellular tissue models, as well as to methods for preparing these cells. The present invention also pertains to a cell fraction comprising liver progenitor cells, liver sinusoidal endothelial progenitor cells, or hepatic stellate progenitor cells. The present invention also pertains to a pharmaceutical composition or kit comprising the above-mentioned cells, a liver cellular tissue model, or a cell fraction. The present invention also pertains to: a method for screening liver disease treatment agents; a method for evaluating the hepatotoxicity of drugs, hepatocytes for infectious disease models, and a method for preparing the same; infectious disease model tissues and a method for preparing the same; as well as a method for screening infectious liver disease treatment agents. 119.-. (canceled)20. A method of preparing hepatic stellate progenitor cells , comprising a step of separating off hepatic stellate progenitor cells from a cell fraction that comprises hepatic stellate progenitor cells , by using the ALCAM-positive phenotype as the marker.21. The method according to claim 20 , further comprising a step of inducing differentiation of pluripotent stem cells to prepare a cell fraction that comprises hepatic stellate progenitor cells.22. The method according to claim 21 , wherein the pluripotent stem cells are human iPS cells.23. Hepatic stellate progenitor cells that can be prepared by the method according to claim 20 , having the ALCAM-positive phenotype claim 20 , having proliferation potency claim 20 , and having differentiation potency to hepatic stellate cells.24. A cell fraction that comprises hepatic stellate progenitor cells at 90% or greater with respect to the total cells claim 20 , wherein the hepatic stellate ...

IN VITRO BILIARY EXCRETION ASSAY

An in vitro methods of characterizing biliary excretion of a chemical entity using a single hepatocyte culture. Comprising providing cell culture comprising hepatocytes forming at least one bile canaliculus; contacting the cell culture with a first chemical entity for a time sufficient to allow uptake of the chemical entity by hepatocytes in the culture; disrupting the at least one bile canaliculus without lysing the hepatocytes and detecting the amount (if any) of the first chemical entity and/or a metabolite thereof released by the at least one bile canaliculus; and lysing the hepatocytes and detecting the amount of the first chemical entity and/or a metabolite thereof released by the hepatocytes. 1. An in vitro method of characterizing biliary excretion of a chemical entity , comprising:a) providing cell culture comprising hepatocytes forming at least one bile canaliculus;b) contacting the cell culture with a first chemical entity for a time sufficient to allow uptake of the chemical entity by hepatocytes in the culture;c) disrupting the at least one bile canaliculus without lysing the hepatocytes and detecting the amount (if any) of the first chemical entity and/or a metabolite thereof released by the at least one bile canaliculus; andd) lysing the hepatocytes and detecting the amount of the first chemical entity and/or a metabolite thereof released by the hepatocytes.2. The method of claim 1 , wherein the cell culture is a hepatocyte-stromal cell coculture comprising hepatocytes and stromal cells disposed on a surface of a solid substrate.3. The method of claim 1 , wherein the amount of the first chemical entity and/or a metabolite thereof released by the at least one bile canaliculus in step c) is higher than the amount of the first chemical entity and/or a metabolite thereof released by the hepatocytes in step d).4. The method of claim 1 , wherein the amount of the first chemical entity and/or a metabolite thereof released by the at least one bile canaliculus ...

Human liver tumor cell line and method of agent screening

An isolated human liver tumor cell line is provided and is named as ITRI-H16, and which was deposited in the Food Industry Research and Development Institute with the accession number BCRC960432 on Nov. 7, 2011. A method of an agent screening is also provided. The method includes providing the isolated human liver tumor cell line ITRI-H16 and treating an interest compound to the ITRI-H16 cell line to determine an effect of the interesting compound on the ITRI-H16 cell line.

Lipopeptides for Use in Treating Liver Diseases and Cardiovascular Diseases

The present invention relates to lipopeptide-based compounds for use in the diagnosis, prevention and/or treatment of a liver disease or condition, preferably liver involved metabolic diseases, as well as in the control or modification of the cholesterol level or cholesterol uptake and, thus, diagnosis, prevention and/or treatment of a cardiovascular disease. The present invention furthermore relates to an in vitro or in vivo assay or method for testing or measuring the NTCP-mediated transport of test compound(s). The present invention furthermore relates to a method for the diagnosis, prevention and/or treatment of a liver disease or condition, comprising administering a therapeutically effective amount of a lipopeptide-based compound to a patient. The present invention furthermore relates to a method for the diagnosis, prevention and/or treatment of a cardiovascular disease. 1. A method for the diagnosis , prevention and/or treatment of a liver disease or condition , wherein said liver disease or condition is related to sodium taurocholate cotransporter polypeptide (NTCP)-mediated transport of compounds into hepatocytes , wherein said method comprises utilizing a lipopeptide-based compound.2. The method of claim 1 , wherein said liver disease or condition that is related to NTCP-mediated transport of compounds into hepatocytes claim 1 , is a liver involved metabolic disease selected from:intrahepatic cholestasis,poisoning of the liver (by liver toxins)/hepatotoxicity,drug-induced cholestatic liver disease,hyperlipidemia, andposthepatic cholestasis.3. The method of claim 1 , wherein the compounds that are transported into hepatocytes via NTCP are:bile acids,taurine- or glycine conjugated bile acids and salts thereof,taurine- or glycine conjugated dihydroxy and trihydroxy bile salts, steroides,', 'steroide sulfates,', 'estrogen conjugates,', 'dehydroepiandrosterone sulfate,', 'conjugated and non-conjugated thyroid hormones,', 'liver toxins.', 'compounds that are ...

HEPATOCYTE PRODUCTION VIA FORWARD PROGRAMMING BY COMBINED GENETIC AND CHEMICAL ENGINEERING

The present invention provides methods comprising both genetic and chemical means for the production of hepatocytes from a variety of cell sources, particularly pluripotent stem cells. 1. A method of producing hepatocytes by forward programming of stem cells , comprising transfecting the stem cells with at least one exogenous expression cassette comprising the hepatocyte programming factor genes encoding FOXA2 , GATA4 , HHEX , HNF1A , and TBX3 , thereby producing hepatocytes from forward programming of the stem cells.2. The method of claim 1 , wherein the at least one exogenous expression cassette is operably linked to an externally inducible transcriptional regulatory element.3. The method of claim 1 , further comprising contacting the stem cells with a MEK inhibitor and/or an ALK5 inhibitor.4. The method of claim 3 , wherein the MEK inhibitor is PD0325901.5. The method of claim 3 , wherein the ALK5 inhibitor is A 83-01.6. The method of claim 3 , further comprising contacting the stem cells with a cyclic AMP analog.7. The method of claim 6 , wherein the cyclic AMP analog is 8-Br-cAMP.8. The method of claim 1 , wherein the stem cells are mesenchymal stem cells claim 1 , hematopoietic stem cells claim 1 , embryonic stem cells claim 1 , or induced pluripotent stem cells.9. The method of claim 1 , wherein the stem cells or progeny cells thereof further comprise a reporter expression cassette comprising a hepatocyte specific transcriptional regulatory element operably linked to a reporter gene.10. The method of claim 9 , wherein the hepatocyte-specific transcriptional regulatory element is a promoter of albumin claim 9 , α-1-antitrypsin (AAT) claim 9 , cytochrome p450 3A4 (CYP3A4) claim 9 , apolipoprotein A-I claim 9 , or APOE.11. The method of claim 1 , wherein the hepatocytes comprise one or more of the hepatocyte characteristics comprising:(i) expression of one or more hepatocyte markers including glucose-6-phosphatase, albumin, α-1-antitrypsin (AAT), cytokeratin 8 ( ...

METHOD OF CULTURING PROLIFERATIVE HEPATOCYTES

A method of culturing animal cells, preferably primary hepatocytes, including a first step of culturing the animal cells in non-adherent culture vessel, preferably a low or ultra-low attachment culture vessel, a second step of embedding the animal cells in a collagen matrix or in a gelatin matrix, and a third step of culturing the animal cells embedded in the collagen matrix or in the gelatin matrix, thereby obtaining 3D animal cell structures including proliferative animal cells, preferably spheroids including proliferative primary hepatocytes. Also, a spheroid including proliferative primary hepatocytes and the uses thereof for engineering an artificial liver model or an artificial liver organ, and for assessing in vitro the liver toxicity, genotoxicity and/or the effects of a drug or a compound. 115-. (canceled)16. A method of culturing animal cells to obtain 3D animal cell structures comprising proliferative animal cells , said method comprising:a) first culturing the animal cells in a non-adherent culture vessel;b) then transferring the animal cells to a culture medium comprising collagen, or truncated collagen also known as gelatin, thereby embedding the animal cells in a collagen or gelatin matrix; andc) culturing the animal cells embedded in the collagen or gelatin matrix;thereby obtaining 3D animal cell structures comprising proliferative animal cells.17. The method according to claim 16 , said method comprising:a) first culturing the animal cells in a non-adherent culture vessel;b) then transferring the animal cells to a culture medium comprising collagen, thereby embedding the animal cells in a collagen matrix; andc) culturing the animal cells embedded in the collagen matrix;thereby obtaining 3D animal cell structures comprising proliferative animal cells.18. The method according to claim 17 , wherein at step b) the animal cells are transferred to a culture medium comprising collagen at a concentration ranging from about 0.25 mg/mL to about 3 mg/mL.19. ...

METHOD FOR MEASURING CELLULAR UPTAKE OF MOLECULES

The present invention provides a method for measuring a cellular uptake amount of a molecule, comprising (i) adding the molecule to an organ-derived cell population to perform incubation, (ii) sorting the organ-derived cell population based on the expression levels of CD31 and CD45, and (iii) after steps (i) and (ii), measuring the amount of the molecule incorporated into the cell population sorted in the step (ii), wherein the molecule is incorporated into cells via a cell surface receptor. 2. The method according to claim 1 , wherein the method comprises step (ii) subsequent to step (i) claim 1 , and the organ-derived cell population incubated with the added molecule is sorted based on the expression levels of CD31 and CD45.3. The method according to claim 1 , wherein the method comprises step (ii) prior to step (i) claim 1 , and the molecule is added to the organ-derived cell population sorted based on the expression levels of CD31 and CD45 to perform incubation.4. The method according to claim 1 , wherein the molecule is an immune complex or an antibody claim 1 , and the receptor is an Fc receptor.5. The method according to claim 1 , wherein the molecule is an anti-IL-6R antibody claim 1 , and the receptor is IL-6R.6. The method according to claim 1 , wherein the molecule is a nucleic acid claim 1 , and the receptor is Stabilin.7. The method according to claim 1 , wherein the organ-derived cell population is a hepatic nonparenchymal cell population.8. The method according to claim 7 , wherein the hepatic nonparenchymal cell population is a human hepatic nonparenchymal cell population claim 7 , and a CD31CD45cell population is sorted in step (ii).9. The method according to claim 8 , wherein the CD31CD45cell population is a cell population having a fluorescence intensity of CD31 ranging from 400 to 7000 and a fluorescence intensity of CD45 ranging from 100 to 4000 in the detection of CD31 and CD45 expressions by flow cytometry.10. A composition for an uptake assay ...

HUMAN HEPATIC 3D CO-CULTURE MODEL AND USES THEREOF

The invention in general relates to human hepatic 3D co-culture models, more in particular 3D spheroid co-cultures of human hepatocyte-like cells and hepatic stellate cells. Furthermore, the invention provides a method for obtaining such co-cultures, as well as the use of said co-cultures in the identification of pro-fibrotic and/or anti-fibrotic compounds. 1. A 3D co-culture comprising hepatocyte-like cells and hepatic stellate cells (HSCs); wherein said hepatic stellate cells are present in equal or excess amounts of said hepatocyte-like cells.2. The 3D co-culture according to claim 1 , wherein said hepatocyte-like cells are human hepatocyte-like cells selected from primary human hepatocytes claim 1 , HepaRG cells claim 1 , human embryonic stem cells (hESC) differentiated into hepatocyte-like cells claim 1 , human induced pluripotent stem cells (hiPSC) differentiated into hepatocyte-like cells claim 1 , primary fibroblast transdifferentiated into hepatocyte-like cells claim 1 , and hepatocyte-like cell lines.3. The 3D co-culture according to claim 1 , wherein said hepatic stellate cells are human hepatic stellate cells selected from primary human hepatic stellate cells claim 1 , human embryonic stem cells (hESC) differentiated into HSC-like cells claim 1 , human induced pluripotent stem cells (hiPSC) differentiated into HSC-like cells claim 1 , and human hepatic stellate cell lines.4. The 3D co-culture according to claim 1 , wherein said 3D co-culture is free-floating in the culture medium.5. The 3D co-culture according to claim 1 , wherein the amount of HSC cells is about double the amount of hepatocyte-like cells.6. (canceled)8. The screening assay according to claim 7 , wherein Col1a1 claim 7 , Col3a1 and/or Loxl2 expression levels are used as a read-out for hepatic stellate cell activation or transdifferentiation in (c).9. The screening assay according to claim 8 , wherein a compound that increases Col1a1 claim 8 , Col3a1 and/or Loxl2 expression is a pro- ...

CULTURE MEDIUM COMPOSITION AND METHOD OF CULTURING CELL OR TISSUE USING THEREOF

The present invention provides a culture method of cells and/or tissues including culturing cells and/or tissues in a suspended state by using a medium composition wherein indeterminate structures are formed in a liquid medium, the structures are uniformly dispersed in the solution and substantially retain the cells and/or tissues without substantially increasing the viscosity of the solution, thus affording an effect of preventing sedimentation thereof, and the like 1. A medium composition comprising (i) deacylated gellan gum or a salt thereof , and (ii) a polysaccharide other than deacylated gellan gum or a salt thereof , wherein the polysaccharide other than deacylated gellan gum or a salt thereof is at least one kind selected from the group consisting of xanthan gum , alginic acid , carageenan , diutan gum , and a salt thereof.2. The medium composition according to claim 1 , wherein the polysaccharide other than deacylated gellan gum or a salt thereof is alginic acid or a salt thereof.3. The medium composition according to claim 1 , wherein the deacylated gellan gum or salt thereof and the polysaccharide other than deacylated gellan gum or a salt thereof are present at a concentration which allows cells or a tissue to be cultured in suspension standing culture.4. The medium composition according to claim 1 , wherein the composition has a viscosity of not more than 8 mPa·s at 37° C.5. The medium composition according to claim 1 , wherein the polysaccharide other than deacylated gellan gum or a salt thereof is at least one kind selected from the group consisting of methylcellulose claim 1 , locust bean gum claim 1 , and a salt thereof.6. The medium composition according to claim 5 , wherein the polysaccharide other than deacylated gellan gum or a salt thereof is methylcellulose or a salt thereof.7. The medium composition according to claim 5 , wherein the deacylated gellan gum or salt thereof and the polysaccharide other than deacylated gellan gum or a salt ...

HUMAN FIBROLAMELLAR HEPATOCELLULAR CARCINOMAS (hFL-HCCS)

The present disclosure provides a model of human fibrolamellar hepatocellular carcinoma (FL-HCC) cells maintained as a transplantable tumor line in a host and a method to establish a transplantable human FL-HCC tumor line. Methods of ex vivo cultures of the FL-HCC are provided. Methods of diagnosing and treating FL-HCC tumors are also provided. 1. A transplantable tumor line of human fibrolamellar hepatocellular carcinoma (hFL-HCC) cells maintained in a non-human animal.2. The transplantable tumor line of claim 1 , wherein the non-human animal is a NOD scid gamma (NSG) mouse.3. The transplantable tumor line of claim 1 , wherein the hFL-HCC cells are derived from a tumor removed from the liver claim 1 , from the biliary tree claim 1 , from a subcutaneous tumor or from an intraperitoneal (ascites) tumor.4. The transplantable tumor line of claim 1 , wherein the tumor line comprises hFL-HCC cells and mesenchymal cells of the non-human animal.5. The transplantable tumor line of claim 1 , wherein at least 50% of the hFL-HCC cells in the transplantable tumor are cancer stem cells.6. The transplantable tumor line of claim 1 , wherein the hFL-HCC cells express the fusion transcript DNAJB1-PRKACA.7. The transplantable tumor line of claim 1 , wherein the hFL-HCC cells overexpress at least one C10orf128 claim 1 , CA12 claim 1 , CREB3L1 claim 1 , GALNTL6 claim 1 , IRF4 claim 1 , ITPRIP claim 1 , KCNE4 claim 1 , NOVA1 claim 1 , OAT claim 1 , PAK3 claim 1 , PCSK1 claim 1 , PHACTR2 claim 1 , RPS6KA2 claim 1 , SLC16A14 claim 1 , TMEM163 claim 1 , or TNRC6C relative to a control sample.8. The transplantable tumor line of claim 7 , wherein the control sample is selected from the group consisting of hepatocellular carcinomas (HCCs) claim 7 , hepatoblastomas claim 7 , cholangiocarcinomas (CCAs) claim 7 , pancreatic cancer claim 7 , biliary tree stem cells claim 7 , hepatic stem cells claim 7 , hepatoblasts claim 7 , pancreatic stem cells claim 7 , hepatic claim 7 , pancreatic committed ...

MICROWELL PLATE

A microwell plate includes a plate body having a surface on which wells are formed. Each of the wells has an open end, a bottom end and an interior space extended from the open end to the bottom end. Each of the wells has, in the interior space, an opening-side portion, a bottom-side portion and a transition portion which is continuously connected to the opening-side portion and to the bottom-side portion, the opening-side portion has a polygonal cross section in parallel to the surface of the plate body, and the bottom-side portion has cross sections in parallel to the surface which have areas that gradually decrease toward the bottom end. 1. A microwell plate , comprising:a plate body having a surface on which a plurality of wells are formed, each of the wells having an open end, a bottom end and an interior space extended from the open end to the bottom end,wherein each of the wells has, in the interior space, an opening-side portion, a bottom-side portion and a transition portion which is continuously connected to the opening-side portion and to the bottom-side portion, the opening-side portion has a polygonal cross section in parallel to the surface of the plate body, and the bottom-side portion has cross sections in parallel to the surface which have areas that gradually decrease toward the bottom end.2. The microwell plate of claim 1 , wherein the opening-side portion has a first depth claim 1 , the transition portion has a second depth claim 1 , the bottom-side portion has a third depth claim 1 , and each of the wells is formed such that a ratio of the first depth to a sum of the second depth and the third depth is from 3:1 to 4:1.3. The microwell plate of claim 1 , wherein the polygonal cross section of the opening-side portion is in a shape of one of a quadrangle claim 1 , a pentagon and a hexagon.4. The microwell plate of claim 1 , wherein the cross sections of the bottom-side portion have circular shapes.5. The microwell plate of claim 1 , wherein the ...

Methods for Producing Cells Having a Phenotype of a Primary Human Hepatocytes and Compositions

The present disclosure provides methods and compositions relating to in vitro cultures of human hepatocyte cell lines which exhibit a primary human hepatocyte phenotype. Such cell lines are susceptible to infection by a hepatotrophic virus, such as HCV or HBV, and support both viral replication and high levels of viral particle production. Such in vitro cultures find use in production and study of hepatotrophic virus, as well as methods of screening (e.g., for antiviral drugs, assessing drug metabolism), and study of primary human hepatocytes. 1. A method of producing a cell culture comprising cells having a primary human hepatocyte phenotype , the method comprising:culturing a human hepatocellular carcinoma (hHCC) cell line in a culture medium comprising human serum for more than 11 dayswherein said culturing induces differentiation of the hHCC cell line into a cell having a primary human hepatocyte phenotype.2. The method of claim 1 , wherein the culture medium comprises about 1% to 20% human serum.3. The method of claim 1 , wherein the culture medium comprises about 2% to 10% human serum.43. The method of any one of - claims 1 , wherein the hHCC cell line is a HuH-7 or HuH-7-derived cell line.54. The method of any one of - claims 1 , wherein said culturing is without subculturing after 10 days of culturing in the culture medium comprising human serum.65. A cell culture comprising cells produced by the method of any one of -.7. A cell culture comprising:cells having a phenotype of a human primary hepatocyte, wherein the cells are the differentiated progeny of an hHCC cell line; anda culture medium comprising human serum.8. A method for assessing an effect of a candidate agent on a cell having a phenotype of a human primary hepatocyte claims 1 , the method comprising:{'claim-ref': {'@idref': 'CLM-00007', 'claim 7'}, 'contacting the cell culture of with a candidate agent; and'}assaying for the presence of absence of an effect of the candidate agent on a phenotype of ...

RECOMBINANT CELLS AND METHODS OF USING SUCH CELLS TO IDENTIFY CIRCADIAN RHYTHM MODULATORS

The invention provides recombinant cells comprising detectable reporters useful in identifying agents, genes, and other modulators of circadian period length and amplitude. Such modulators are useful for resetting the circadian clock in a variety of contexts (e.g., jet lag, shift work). Such cells are also useful in selecting an administration regimen for a therapeutic agent, where the agent's efficacy and/or adverse side effects show circadian effects. 1. A recombinant cell comprising an expression vector , wherein the expression vector comprises a promoter selected from the group consisting of Period2 (Per2) , Cry1 , Cry1-Intron , and Bmal1 , wherein the promoter is operationally linked to a detectable reporter that is expressed at high-amplitude and with a persistent rhythm.2. A recombinant adipocyte or hepatocyte cell or progenitor thereof comprising an expression vector , wherein the expression vector comprises Period2 (Per2) , Cry1 , Cry1-Intron , and Bmal1 promoter operationally linked to a detectable reporter.3. The recombinant cell of or , wherein the cell is a 3T3-L1 pre-adipocyte or a MMH-D3 pre-hepatocyte.4. The recombinant cell of or , wherein the expression vector is a lentiviral vector.5. The recombinant cell of or , wherein the detectable reporter is a luciferase reporter.6. The recombinant cell of or , wherein the reporter expression varies at least about two to four fold in trough to peak levels.7. The recombinant cell of or , wherein the reporter expression varies at least about three fold in trough to peak levels.8. A method of identifying a circadian cycle modulator , the method comprising contacting the cell of any of - with an agent , and assaying reporter expression in the contacted cell relative to a corresponding control cell.9. The method of claim 8 , wherein the agent is a small compound claim 8 , inhibitory nucleic acid claim 8 , or polypeptide.10. A method of identifying a circadian cycle modulator claim 8 , the method comprising ...

Methods of Using Compositions Comprising Variants and Fusions of FGF19 Polypeptides for Modulating Bile Acid Homeostasis in a Subject Having Cirrhosis

The invention relates to variants and fusions of fibroblast growth factor 19 (FGF19), variants and fusions of fibroblast growth factor 21 (FGF21), fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21), and variants or fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21) proteins and peptide sequences (and peptidomimetics), having one or more activities, such as bile acid homeostasis modulating activity, and methods for and uses in treatment of bile acid and other disorders. 171-. (canceled)72. A method of reducing bile acid synthesis in a subject having cirrhosis , comprising administering to the subject an effective amount of a peptide , wherein the peptide comprises:a) an N-terminal region comprising at least seven amino acid residues, the N-terminal region having a first amino acid position and a last amino acid position, wherein the N-terminal region comprises DSSPL (SEQ ID NO: 121) or DASPH (SEQ ID NO: 122); and (i) a first C-terminal region sequence comprising WGDPIRLRHLYTSG (amino acids 16 to 29 of SEQ ID NO:99 [FGF19]), wherein the W residue corresponds to the first amino acid position of the C-terminal region; and', '(ii) a second C-terminal region sequence comprising PHGLSSCFLRIRADGVVDCARGQSAHSLLEIKAVALRTVAIKGVHS VRYLCMGADGKMQGLLQYSEEDCAFEEEIRPDGYNVYRSEKHRL PVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESD MFSSPLETDSMDPFGLVTGLEAVRSPSFEK (amino acid residues 30 to 194 of SEQ ID NO:99 [FGF19]); or a sequence comprising from 1 to 5 amino acid substitutions, deletions or insertions thereof,, 'b) a C-terminal region comprising a portion of SEQ ID NO:99 [FGF19], the C-terminal region having a first amino acid position and a last amino acid position, wherein the C-terminal region comprises'}wherein the peptide(A) has reduced hepatocellular carcinoma (HCC) formation as compared to FGF 19, or as compared to an FGF 19 variant sequence having any of GQV, GDI, WGPI (SEQ ID NO:171), WGDPV (SEQ ID NO: ...

Methods of Using Compositions Comprising Variants and Fusions of FGF19 Polypeptides for Modulating Bile Acid Homeostasis in a Subject Having Liver Fibrosis

The invention relates to variants and fusions of fibroblast growth factor 19 (FGF19), variants and fusions of fibroblast growth factor 21 (FGF21), fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21), and variants or fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21) proteins and peptide sequences (and peptidomimetics), having one or more activities, such as bile acid homeostasis modulating activity, and methods for and uses in treatment of bile acid and other disorders. 171-. (canceled)72. A method of reducing bile acid synthesis in a subject having liver fibrosis , comprising administering to the subject an effective amount of a peptide , wherein the peptide comprises:a) an N-terminal region comprising at least seven amino acid residues, the N-terminal region having a first amino acid position and a last amino acid position, wherein the N-terminal region comprises DSSPL (SEQ ID NO:121) or DASPH (SEQ ID NO:122); and (i) a first C-terminal region sequence comprising WGDPIRLRHLYTSG (amino acids 16 to 29 of SEQ ID NO:99 [FGF19]), wherein the W residue corresponds to the first amino acid position of the C-terminal region; and', '(ii) a second C-terminal region sequence comprising PHGLSSCFLRIRADGVVDCARGQSAHSLLEIKAVALRTVAIKGVHS VRYLCMGADGKMQGLLQYSEEDCAFEEEIRPDGYNVYRSEKHRL PVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESD MFSSPLETDSMDPFGLVTGLEAVRSPSFEK (amino acid residues 30 to 194 of SEQ ID NO:99 [FGF19]); or a sequence comprising from 1 to 5 amino acid substitutions, deletions or insertions thereof;, 'b) a C-terminal region comprising a portion of SEQ ID NO:99 [FGF19], the C-terminal region having a first amino acid position and a last amino acid position, wherein the C-terminal region comprises'}wherein the peptide(A) has reduced hepatocellular carcinoma (HCC) formation as compared to FGF19, or as compared to an FGF19 variant sequence having any of GQV, GDI, WGPI (SEQ ID NO:171), WGDPV (SEQ ID NO: ...

Methods of Using Compositions Comprising Variants and Fusions of FGF19 Polypeptides for Modulating Bile Acid Homeostasis in a Subject Having Nonalcoholic Fatty Liver Disease

The invention relates to variants and fusions of fibroblast growth factor 19 (FGF19), variants and fusions of fibroblast growth factor 21 (FGF21), fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21), and variants or fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21) proteins and peptide sequences (and peptidomimetics), having one or more activities, such as bile acid homeostasis modulating activity, and methods for and uses in treatment of bile acid and other disorders.

Induced Hepatocytes and Uses Thereof

Disclosed herein are induced hepatocytes from a trophoblast stem cell, methods for inducing the cells, and compositions thereof. Also disclosed herein are methods of treating a disease or disorder (e.g., liver-associated) by utilizing an induced hepatocyte disclosed herein. 1148-. (canceled)149. A method of inducing a trophoblast stem (TS) cell to differentiate into an induced hepatocyte in vitro , comprising:contacting the trophoblast stem cell with a conditioned medium to induce differentiation of the trophoblast stem cell into an induced hepatocyte,wherein the conditioned medium comprises a fibroblast growth factor (FGF), a steroid, and a cytokine.150. The method of claim 149 , wherein the induced hepatocyte is a hepatic progenitor cell.151. The method of claim 149 , wherein the TS cell is a human TS cell.152. An induced hepatocyte produced by the method of .153. An isolated induced hepatocyte derived from a trophoblast stem cell claim 149 , wherein the hepatocyte expresses one or more biomarkers selected from the group consisting of human leukocyte antigen G (HLA-G) claim 149 , transforming growth factor beta 1 (TGFβ1) claim 149 , cluster of differentiation 4 (CD4) claim 149 , forkhead box P3 (Foxp3) claim 149 , human cytoplasmic marker stem 121 (stem 121) claim 149 , mast/stem cell growth factor receptor C-kit (C-kit) claim 149 , betatrophin claim 149 , apolipoprotein F (APOF) claim 149 , alcohol dehydrogenase-1 (ADH1) claim 149 , carbamoyl-phosphate synthase 1 (CPS1) claim 149 , GATA transcription factor 4 (GATA4) claim 149 , cytochrome P450 family 1 subfamily A polypeptide 1 (CYP1A1) claim 149 , cytochrome P450 2B6 (CYP2B6) claim 149 , asialoglycoprotein receptor 1 (ASGR1) claim 149 , C-X-C chemokine receptor type 4 (CXCR4) claim 149 , bile salt export pump (BSEP) claim 149 , multi-drug resistance protein-2 (MRP2) claim 149 , connexin 32 (CX32) claim 149 , forkhead box protein A2 (FOXA2) claim 149 , SRY-box 17 (SOX17) claim 149 , hexosaminidase A alpha ...

Human Hepatoma Cell Line HLCZ01 and Uses Thereof

A human hepatoma cell line is HLCZ01 cell line which has been deposited in the China Center for Type Culture Collection (CCTCC), and having Accession (deposit) No. is CCTCC NO: C201309. The human hepatoma cell line HLCZ01 is used as a cell model supporting said hepatitis viruses infection and is used to establish an animal model for supporting virus infection, wherein said human hepatoma cell line HLCZ01 is also used for preparation, screening and evaluating anti-hepatitis virus drugs, anti-tumor drugs, and used for manufacturing artificial liver. 110-. (canceled)11. A hepatoma cell line , which comprises a human hepatoma cell line HLCZ01 having 54 to 63 chromosomes.12. The hepatoma cell line claim 11 , as recited in claim 11 , wherein said human hepatoma cell line HLCZ01 shows a phenomenon of heteromorphic chromosomes after karyotype analysis of chromosome G band claim 11 , and a marker chromosome appears in a karyotype.13. The hepatoma cell line claim 11 , as recited in claim 11 , wherein said human hepatoma cell line HLCZ01 comprises hepatocyte specific genes ALB and AAT.14. The hepatoma cell line claim 12 , as recited in claim 12 , wherein said human hepatoma cell line HLCZ01 comprises hepatocyte specific genes ALB and AAT.15. The hepatoma cell line claim 14 , as recited in claim 14 , wherein said hepatocyte specific genes ALB and AAT are detected by RT-PCR.16. The hepatoma cell line claim 11 , as recited in claim 11 , wherein said human hepatoma cell line HLCZ01 expresses liver cell specific protein ALB and AAT.17. The hepatoma cell line claim 12 , as recited in claim 12 , wherein said human hepatoma cell line HLCZ01 expresses liver cell specific protein ALB and AAT.18. The hepatoma cell line claim 17 , as recited in claim 17 , wherein said liver cell specific protein ALB and AAT are detected by a Western Blot process.19. The hepatoma cell line claim 11 , as recited in claim 11 , wherein said human hepatoma cell line HLCZ01 is infectable by a hepatitis virus.10 ...

Hepatocytes and hepatic non-parenchymal cells, and methods for preparation thereof

The present invention pertains to hepatocytes, liver progenitor cells, cholangiocytes, liver sinusoidal endothelial progenitor cells, liver sinusoidal endothelial cells, hepatic stellate progenitor cells, hepatic stellate cells, and liver cellular tissue models, as well as to methods for preparing these cells. The present invention also pertains to a cell fraction comprising liver progenitor cells, liver sinusoidal endothelial progenitor cells, or hepatic stellate progenitor cells. The present invention also pertains to a pharmaceutical composition or kit comprising the above-mentioned cells, a liver cellular tissue model, or a cell fraction. The present invention also pertains to: a method for screening liver disease treatment agents; a method for evaluating the hepatotoxicity of drugs, hepatocytes for infectious disease models, and a method for preparing the same; infectious disease model tissues and a method for preparing the same; as well as a method for screening infectious liver disease treatment agents.

Componential Analyzer, Drug Efficacy Analyzer, and Analysis Method

Application of the present invention enables quantification of fractions of candidate pharmaceutical compounds (a parent compound and its metabolites), one excreted to the basolateral (Basal/Basolateral)-side via transporters and by diffusion, one excreted to the lumen (Apical)-side, and one remained in the cells. This enables determination of the total amount of the administered candidate pharmaceutical compounds and the distribution ratio of the fractions. The kinetics of the administered candidate pharmaceutical compounds can be evaluated, thereby enabling in vitro screening of an enormous number of candidate pharmaceutical compounds for drug candidates exhibiting the efficacy. The object of the present invention is to provide an apparatus and method for understanding a total picture of pharmacokinetics in vitro by quantifying a fraction of basolateral (Basal/Basolateral) efflux, a fraction of lumen (Apical)-side excretion, and a fraction remaining in a cell of a drug which has been administered to the cell to determine the distribution ratio of each fraction. 1. A componential analyzer , comprising:a holding unit for holding a plurality of containers holding a predetermined cell;a temperature controlling unit for controlling temperatures in the plurality of containers; andan analyzing unit for measuring a component in the plurality of containers and analyzing the measured component,wherein the plurality of containers comprises at least a first container and a second container;the first container and the second container each contain a first buffer solution;the temperature controlling unit controls temperature in the first container and temperature in the second container so that the temperatures are different from each other; andthe analyzing unit measures:an amount of the component excreted from a cell in the first container to the first buffer solution in the first container; andan amount of the component excreted from a cell in the second container to the ...

COMPOSITIONS AND METHODS FOR IDENTIFYING AND TREATING CACHEXIA OR PRE-CACHEXIA

The invention provides markers indicative of pre-cachexia, compositions and methods for identifying patients with a molecular signature indicative of pre-cachexia; a culture system that reproduces the cachetic process in cells in vitro, which facilitates the screening and identification of therapeutic agents useful for disrupting (slowing, reducing, reversing, or preventing) the progression of pre-cachexia to refractory cachexia; as well as therapeutic agents identified using the culture system of the invention. 185.-. (canceled)86. A method of inhibiting the loss of myosin heavy chain in a myocyte , inhibiting lipolysis in an adipocyte , or inhibiting atrophy in a myocyte , adipocyte , or hepatocyte , the method comprising:contacting the myocyte, adipocyte, or hepatocyte with an effective amount of an agent that inhibits Receptor for Advanced Glycation Endproducts (RAGE) activity, wherein the RAGE inhibitor agent is recombinant soluble RAGE (rsRAGE), an anti-RAGE antibody, a RAGE blocking peptide that inhibits binding of RAGE to ligands, a small molecule that inhibits binding of RAGE to ligands, a dominant negative RAGE, an inhibitor of RAGE phosphorylation, an inhibitor of RAGE aggregation, or an inhibitor of RAGE interactions with other heterotypic receptors.87. The method of claim 86 , wherein the myocyte claim 86 , adipocyte claim 86 , or hepatocyte cell is in vitro or in vivo.88. The method of claim 86 , wherein the agent that inhibits RAGE activity is PF-04494700 claim 86 , FPS1 claim 86 , FPS2 claim 86 , FPS3 or FPS-ZM1.89. The method of claim 87 , wherein the myocyte claim 87 , adipocyte claim 87 , or hepatocyte cell is present in a subject identified as having at least one cancer selected from the group consisting of carcinomas claim 87 , sarcomas claim 87 , cancers of the skin claim 87 , pancreas claim 87 , stomach claim 87 , colon claim 87 , thorax claim 87 , liver claim 87 , gallbladder claim 87 , musculoskeletal system claim 87 , breast claim 87 , lung ...

Screening method for selected amino lipid-containing compositions

The invention features a method of identifying therapeutically relevant compositions which include a therapeutic agent and 2,2-Dilinoley 1-4-dimethylaminomethyl-[1,3]-dioxolane by screening for an effect of the agent on the liver of a model subject.

Utility of protein in the prediction of in vivo effects

A method of evaluating disposition and/or effect of a candidate compound in an in vitro culture and/or suspension to predict in vivo disposition and/or effect of the candidate compound, which can include the steps of providing a cell culture and/or suspension; exposing a candidate compound to the culture and/or suspension; exposing the culture and/or suspension to a media providing an in vivo relevant extracellular environment, such as a media comprising a component (such as a protein and/or other component, such as lipoproteins, bile acids, and endogenous compounds such as bilirubin) at a physiologic concentration or a concentration having characteristics, such as binding characteristics, similar to the physiological concentration; wherein any combination of any of the exposing steps can occur in any order or simultaneously; and evaluating in vitro disposition and/or effect of the compound to predict in vivo disposition and/or effect of the candidate compound.

CELL CULTURE MEDIA FOR DIFFERENTIATION OF STEM CELLS INTO HEPATOCYTES

The invention relates to methods and media for inducing or maintaining a mature hepatocyte phenotype in a cell, the method comprising the step of cultivation cells in a cell culture medium comprising at least 10 mg amino acids/ml medium, wherein at least 50% (w/w) of the concentration of said amino acids in said medium is provided by one up to five amino acids and, with the proviso that said one up to five amino acids is not one of Thr, Tyr, Cys, Met, Arg and His. 1. A method for inducing or maintaining a mature hepatocyte phenotype in a cell , the method comprising cultivating cells in a cell culture medium comprising at least 10 mg amino acids/ml medium ,wherein at least 50% (w/w) of the concentration of the amino acids in the medium is provided by from one to five amino acids and, with the proviso that the from one to five amino acids is not one of Thr, Tyr, Cys, Met, Arg or His.2. The method according to claim 1 , wherein at least 70% (w/w) of the concentration of the amino acids in the medium is provided by the from one to five amino acids.3. The method according to claim 1 , wherein the medium comprises at least 15 mg amino acids/ml medium.4. (canceled)5. The method according to claim 1 , wherein the from one to five amino acids are selected from the group consisting of Gly claim 1 , Ala claim 1 , Ser Val claim 1 , Ile claim 1 , Leu claim 1 , Pro claim 1 , Asp and taurine.6. (canceled)7. The method according to claim 1 , wherein at least 50% (w/w) of the concentration of the amino acids in the medium is Glycine.8. The method according to claim 1 , further comprising determining a marker of mature phenotype selected from the group consisting of albumin secretion claim 1 , CYP450 activity claim 1 , glycogen storage claim 1 , susceptibility to infection with hepatotropic viruses claim 1 , drug metabolising capacities and expression of NTCP claim 1 , HNF4A and AAT.9. (canceled)10. The method according to claim 1 , wherein the cultivation in the medium is performed ...

Method of measuring inhibition of phosphatidylcholine export transport and/or formation activity

A method is provided to measure modulation of phosphatidylcholine export transport and/or formation activity in hepatocyte or stable cell-line preparations by test agents including but not limited to drugs, drug candidates, biologicals, food components, herb or plant components, proteins, peptides, DNA, and RNA. Furthermore, the method is designed to determine modulation of phosphatidylcholine transport and/or formation activity not only by said test agents, but also their metabolites or biotransformed products formed in situ.

Screening method for selected amino-lipid-containing compositions

The invention features a method of identifying therapeutically relevant compositions which include a therapeutic agent and 2,2-dimethylaminomethyl-[1-3]-dioxolane by screening for an effect of the agent on the liver of a model subject.

Hsd17b13 variants and uses thereof

Provided are compositions related to HSD17B13 variants, including nucleic acid molecules and polypeptides related to variants of HSD17B13, and cells comprising those nucleic acid molecules and polypeptides. Also provided are methods related to HSD17B3 variants. Such methods include methods for detecting the presence of the HSD17B13 rs72613567 variant in a biological sample comprising genomic DNA, for detecting the presence or levels of any one of variant HSD17B13 Transcripts C, D, E, F, G, and H, and particularly D, in a biological sample comprising mRNA or cDNA, or for detecting the presence or levels of any one of variant HSD17B13 protein Isoforms C, D, E, F, G, or H, and particularly D, in a biological sample comprising protein. Also provided are methods for determining a subject's susceptibility to developing a liver disease or of diagnosing a subject with liver disease.

Culture medium composition and method of culturing cell or tissue using thereof

The present invention provides a culture method of cells and/or tissues including culturing cells and/or tissues in a suspended state by using a medium composition wherein indeterminate structures are formed in a liquid medium, the structures are uniformly dispersed in the solution and substantially retain the cells and/or tissues without substantially increasing the viscosity of the solution, thus affording an effect of preventing sedimentation thereof, and the like

IN VITRO MODEL FOR PATHOLOGICAL OR PHYSIOLOGIC CONDITIONS

The present invention generally relates to in vitro methods for mimicking in vivo pathological or physiologic conditions. The methods comprise applying shear forces to a cell type or cell type plated on a surface within a cell culture container. Methods for testing drugs or compounds in such systems are also described. 1259-. (canceled)260. A method for mimicking a pathological condition of the liver in vitro , the method comprising:adding a culture medium and serum from a subject comprising at least one factor to a cell culture container;plating hepatocytes on at least one surface within the cell culture container, wherein the hepatocytes comprise hepatocytes isolated from at least one subject having the pathological condition; andapplying a shear force upon the plated hepatocytes, the shear force resulting from flow of the culture medium induced by a flow device, the flow mimicking flow to which the hepatocytes are exposed in vivo in the pathological condition.261. The method of claim 260 , wherein the hepatocytes comprise primary cells.262. The method of claim 260 , wherein the surface within the cell culture container comprises a porous membrane suspended in the cell culture container.263. The method of claim 260 , wherein the shear force is applied indirectly to the plated hepatocytes.264. The method of claim 263 , wherein the hepatocytes are plated on a first surface of a porous membrane claim 263 , the porous membrane is suspended in the cell culture container such that the first surface is proximal and in spaced relation to a bottom surface of the cell culture container claim 263 , thereby defining within the cell culture container a lower volume comprising the hepatocytes and an upper volume comprising a second surface of the porous membrane claim 263 , and the shear force is applied upon the second surface of the porous membrane in the upper volume of the container.265. The method of claim 264 , further comprising plating nonparenchymal hepatic cells on ...

NEXT GENERATION DESIGNER LIVER ORGANOIDS AND THEIR METHODS OF PREPARATION AND USE

The present disclosure relates to synthetic liver organoids and methods of using such synthetic liver organoids for various applications including drug discovery and modeling human liver development. In particular, provided herein are methods of producing and using synthetic mature liver organoids comprising mature, functional cells found in the human liver. 1. A method for producing a synthetic mature liver organoid , the method comprising the steps of{'sup': '+', '(a) introducing into cells of a Day-5 fetal liver organoid one or more lentiviral constructs comprising an inducible transgene encoding at least one transcription factor selected from ATF5, Prox1, MLXIPL1, and CREB3L3, wherein, prior to introducing the one or more lentiviral constructs, the Day-5 fetal liver organoid comprising a cell population comprising at least 70% CXCR4 cells;'}(b) inducing expression of the inducible transgene by contacting the fetal liver organoid of step (a) to a small-molecule inducer of transgene expression;(c) culturing the induced organoid of step (b) for about 5 days;(d) transducing cells of the cultured organoid of step (c) with one or more CRISPR cassettes comprising a nucleic acid sequence encoding dCas9 and one or more gRNAs that bind to the human CYP3A4 locus; and{'sup': +', '+', '+', '+, '(e) culturing the transduced cultured organoids of step (d) for at least 5 days, thereby producing a mature liver organoid comprising albumin/FGF21/G6PC/FXR hepatocytes and exhibiting CYP3A4 protein metabolizing activity, hepatic bile acid receptor FXR activity, and bile acid production.'}2. The method of claim 1 , wherein the transgene is inducible by doxycycline.3. The method of claim 1 , wherein the one or more gRNAs is under control of a constitutively active phEF1a promoter.4. The method of claim 1 , wherein the one or more gRNAs is under control of a pAAT promoter.5. The method of claim 1 , wherein the mature liver organoid exhibits one or more properties selected from the group ...

STEM CELL-DERIVED HEPATOCYTES IN CO-CULTURE AND USES THEREOF

The present disclosure provides co-cultures of human pluripotent stem cell derived hepatocytes and at least one non-parenchymal cell population in vitro, methods of preparing the co-cultures and methods of using the co-cultures for high throughput screening and evaluation of drug candidates. The stem cell derived hepatocyte co-culture system provides an in vitro model in which cell viability and relatively mature hepatocyte phenotype of stem cell derived hepatocytes are maintained for extended periods relative to conventional monoculture. 1. A composition comprising a population of hepatocytes derived from pluripotent stem cells and at least one non-parenchymal cell population in co-culture in vitro , and a layer of material comprising at least one extracellular matrix protein disposed on the co-culture.2. The composition of claim 1 , further comprising a culture substrate claim 1 , wherein the cell populations are disposed in a micropattern on the culture substrate.3. The composition of claim 2 , wherein the culture substrate comprises a glass surface claim 2 , a polystyrene surface claim 2 , or a silicon surface.4. The composition of claim 2 , wherein the micropattern comprises a predetermined two-dimensional pattern of multiple microdots claim 2 , the micropattern defined by a microdot diameter and a center-to-center spacing between each of any two neighboring microdots.5. The composition of claim 4 , wherein each microdot has a diameter of about 500 μm to about 700 μm.6. The composition of claim 4 , wherein the center-to-center spacing between each microdot is about 1200 μm.7. The composition of claim 1 , wherein the hepatocytes are derived from induced pluripotent human stem cells.8. The composition of claim 1 , following co-culture in vitro for a period and under culture conditions sufficient to allow the hepatocytes to maintain a higher level of differentiation toward an adult human hepatocyte phenotype when compared to hepatocytes derived from induced ...

QEEG/Genomic Analysis For Predicting Therapeutic Outcome

Using a combinatorial algorithm comprised of quantitative EEG features and at least one pharmacogenomic variable, a significantly higher predictive accuracy and usability is achieved as compared to other current methods of clinical decision support for guided pharmacotherapy. The method produces a report with actionable findings for the treating physician, recommending for and/or against multiple drug classes and agents from among the available treatments for mental health disorders. While predictive accuracy for pharmacogenomic testing averages 73%, the presently disclosed combinatorial algorithms achieve a significantly higher rate of accuracy at 91%. 1. A method , comprising:a) collecting an electroencephalogram and a plurality of cells from a patient exhibiting at least one symptom of a diagnosed psychiatric disorder;b) converting said electroencephalogram into at least one quantitative electroencephalographic (QEEG) feature variable;c) identifying at least one genotype in said plurality of cells;d) comparing said at least one QEEG feature variable to a first database to create a first therapy list prioritized according to a first predicted efficacy score, said first therapy list comprising a first recommended therapy;e) comparing said at least one genotype to a second database to create a second therapy list prioritized according to a second predicted efficacy score, said second therapy list comprising a second recommended therapy;f) matching said first therapy list and said second therapy list to create a final therapy list prioritized according to a combined first and second efficacy score, said final therapy list comprising a final recommended therapy; andg) administering said final recommended therapy to said patient under conditions such that said at least one symptom is reduced, wherein said selected therapy comprises a combined first and second efficacy score that is within a preferred range.2. The method of claim 1 , wherein said final recommended ...

SCREENING METHOD FOR SELECTED AMINO-LIPID-CONTAINING COMPOSITIONS

The invention features a method of identifying therapeutically relevant compositions which include a therapeutic agent and 2,2-dimethylaminomethyl-[1-3]-dioxolane by screening for an effect of the agent on the liver of a model subject. 1. A method of evaluating a composition that includes a therapeutic agent and 2 ,2-Dilinoley 1-4-dimethylaminomethyl-[1 ,3]-dioxolane comprising:providing a composition that includes a therapeutic agent and 2,2-Dilinoley 1-4-dimethylaminomethyl-[1,3]-dioxolane;administering the composition to a test animal; anddetermining the effect of the composition on the expression of a target gene expressed in the liver of the animal,thereby evaluating the composition.2. The method of claim 1 , wherein the therapeutic agent is an RNA-based construct.3. The method of claim 2 , wherein the RNA-based construct is a dsRNA.4. The method of claim 1 , wherein the target gene is Factor VII.5. The method of claim 1 , wherein determining the effect of the composition comprises determining target protein levels.6. The method of claim 1 , wherein determining the effect of the composition comprises determining target mRNA levels.7. The method of claim 5 , wherein the level of target protein in blood is determined.8. The method of claim 6 , wherein the level of target mRNA in liver is determined.9. The method of claim 1 , further comprising comparing expression of the target gene with a preselected reference value.10. The method of claim 1 , wherein the composition further comprises a third component.11. The method of claim 1 , wherein the therapeutic agent is an antisense RNA claim 1 , ribozyme or microRNA.12. The method of claim 1 , wherein the test animal is a rodent.13. The method of claim 1 , wherein the test animal is a mouse.14. The method of claim 1 , wherein the composition reduces FVII protein or mRNA levels in the blood.15. The method of claim 1 , wherein the composition reduces FVII protein or mRNA levels in the liver.16. The method of claim 1 , ...

IDENTIFYING CANCER THERAPIES

In silico tools are used to determine possibly effective therapies for treating a patient's cancer based on patient, drug, and cancer information. Functional assays can be performed on living cancer cells from the patient to evaluate the possibly effective therapies along with subsequent genomic or other more destructive assays to provide additional information from a single sample. Drug, patient, cancer, and outcome information can be recorded and updated iteratively and analyzed using machine learning to identify correlations between various patient, cancer, and drug characteristics and expected outcomes and drug efficacies. 1. A method for selecting a cancer treatment , the method comprising:obtaining a biological sample containing cancer cells from a patient;identifying, in silico, a therapeutic that meets predetermined criteria relating to one or more selected from the group of toxicology, efficacy, pharmacokinetics, side-effects, drug interactions, patient compliance, and cost;performing an in vitro assay to determine efficacy of the identified therapeutic; andwhere the efficacy is above a threshold, selecting the identified therapeutic for treating cancer in the patient.2. The method of claim 1 , wherein the biological sample is from a patient having received a prior therapy for cancer.3. The method of claim 2 , wherein the predetermined criteria further comprises the prior therapy the patient has received.4. The method of claim 1 , wherein the in vitro assay measures a functional feature of live cells of the biological sample.5. The method of claim 4 , wherein the functional feature comprises change in mass of the live cancer cells.6. The method of claim 5 , wherein the change in mass is measured using at least one suspended microchannel resonator.7. The method of claim 4 , further comprising isolating individual claim 4 , live cells claim 4 , from the biological sample before performing the in vitro assay on the individual claim 4 , live cells.8. The method ...

Metabolic restriction in cell-based assays

Provided are methods and assays used in the screening and identification of agents in assays that use metabolically active cells.

Microscale micropatterned engineered in vitro tissue

The disclosure provides an in vitro culture systems. The invention provides methods and systems useful for developing in vitro an engineered tissue, method of using the tissue and compositions comprising the tissue.

Methods and compositions to treat liver diseases and conditions

Methods and compounds useful to treat diseases and conditions are provided. Methods of administering one or more microRNA-inhibitor compounds or one or more microRNA-enhancing compounds to cells, tissues, and/or subjects as a treatment for a disease or condition are provided.

MICROFLUIDIC DEVICE FOR CELL-BASED ASSAYS

A microfluidic device, method and kit for assaying and/or culturing cells are provided. The microfluidic device comprises a well block comprising a plurality of microwells; at least one cell culture layer selected from a first cell culture layer comprising a plurality of microchannels, each microchannel being aligned with one of the plurality of microwells and being in fluid communication with the aligned microwells; and a second cell culture layer comprising a plurality of cell culture chamber wells, each cell culture chamber well being aligned with one of the plurality of microwells and being in fluid communication with the aligned microwells, and a plurality of outlets, each of the plurality of outlets corresponding to one of the plurality of cell culture chamber wells; and a base block, wherein the at least one cell culture layer is sealably coupled between the well block and the base block, thereby allowing fluid communication between the plurality of microwells in the well block and the at least one cell culture layer. 1. A microfluidic device for assaying cells , the microfluidic device comprising:a well block comprising a plurality of microwells;a first cell culture layer comprising a plurality of microchannels, each microchannel being aligned with one of the plurality of microwells and being in fluid communication with the aligned microwells; anda base block, the base block being in fluid communication with the plurality of microchannels, wherein the first cell culture layer is sealably coupled between the well block and the base block, thereby allowing fluid communication between the plurality of microwells in the well block, the aligned microchannels in the first cell culture layer and the base block.2. The microfluidic device of claim 1 , wherein the plurality of microchannels have defined geometries that produce one or more desired flow rates through the plurality of microchannels.3. (canceled)4. The microfluidic device of claim 1 , wherein an internal ...

USE OF STING AGONISTS TO TREAT HEPATITIS B VIRUS INFECTION

The invention includes methods of treating a subject having hepatitis B viral (HBV) infection. In certain embodiments, the method of the invention comprises stimulating the innate cytokine response in macrophages, dendritic cells and/or liver non-parenchymal cells with small molecular STING agonists, thus suppressing HBV replication in hepatocytes. In other embodiments, the method of the invention can be used to treat chronic HBV infections. The invention further provides methods of identifying compounds useful in treating HBV infection in a subject. 1. A method of treating a subject having a hepatitis B viral (HBV) infection , the method comprising administering a pharmaceutically effective amount of a STING agonist to the subject.2. The method of claim 1 , wherein the STING agonist stimulates an innate cytokine response in at least one cell selected from the group consisting of macrophages claim 1 , dendritic cell claim 1 , liver non-parenchymal cell claim 1 , and any combinations thereof.3. The method of claim 2 , wherein the innate cytokine response is mediated through cytokines.4. The method of claim 3 , wherein the innate cytokine response is mediated through type 1 interferon.5. The method of claim 1 , wherein the STING agonist comprises a flavonoid.6. The method of claim 5 , wherein the flavonoid comprises at least one selected from the group consisting of 10-(carboxymethyl) 9(10H)acridone (CMA) claim 5 , 5 claim 5 ,6-Dimethylxanthenone-4-acetic acid (DMXAA) claim 5 , methoxyvone claim 5 , 6 claim 5 ,4′-dimethoxyflavone claim 5 , 4′-methoxyflavone claim 5 , 3′ claim 5 ,6′-dihydroxyflavone claim 5 , 7 claim 5 ,2′-dihydroxyflavone claim 5 , daidzein claim 5 , formononetin claim 5 , retusin 7-methyl ether claim 5 , xanthone claim 5 , and any combinations thereof.7. The method of claim 6 , wherein the flavonoid comprises DMXAA.8. The method of claim 1 , wherein the STING agonist suppresses hepatitis B virus replication in infected hepatocytes.9. The method of ...

IN VITRO TEST SYSTEM TO EVALUATE XENOBIOTICS AS IMMUNE-MODULATORS OF DRUG TRANSPORT AND METABOLISM IN HUMAN HEPATOCYTES

A kit and method for evaluating in vitro the effect of a xenobiotic (e.g., a biologic drug) on drug metabolism in hepatocytes. The kit comprises a first culture of hepatocytes, a portion of in vitro xenobiotic-stimulated biological sample, and instructions for incubating the first culture of hepatocytes with the portion of in vitro xenobiotic-stimulated biological sample, and analyzing the activity, expression, or a combination thereof of a biomarker in the hepatocytes to evaluate the effect of the xenobiotic on drug metabolism in the hepatocytes. 1. A kit for evaluating in vitro the effect of a xenobiotic on drug metabolism in hepatocytes , said kit comprising:a first culture of hepatocytes;an in vitro xenobiotic-stimulated biological sample; andinstructions for incubating said first culture of hepatocytes with said in vitro xenobiotic-stimulated biological sample and analyzing the activity, expression, or a combination thereof of a biomarker in said hepatocytes to evaluate the effect of said xenobiotic on drug metabolism in said hepatocytes.2. The kit of claim 1 , wherein said hepatocytes are cryopreserved hepatocytes.3. The kit if claim 1 , wherein said first culture of hepatocytes is a co-culture of hepatocytes and Kupffer cells.4. The kit if claim 1 , wherein said hepatocytes are human hepatocytes.5. The kit if claim 1 , wherein said hepatocytes are pooled hepatocytes.6. The kit of claim 1 , wherein said in vitro xenobiotic-stimulated biological sample is a portion of in vitro xenobiotic-stimulated biological sample prepared by exposing a first culture of said biological sample to said xenobiotic in vitro to yield a xenobiotic-stimulated biological sample claim 1 , and separating said portion of said xenobiotic-stimulated biological sample.7. The kit of claim 6 , wherein said portion of in vitro xenobiotic-stimulated biological sample is plasma that has been separated from in vitro xenobiotic-stimulated whole blood.8. The kit of claim 6 , wherein said portion ...

Plated hepatocytes and preparation and uses thereof

The present invention provides a product comprising plated human hepatocytes on a surface and at least some of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, which are attached to the surface. A method of preparing plated human hepatocytes is also provided. The preparation method comprises applying human hepatocytes to a surface in the presence of feeder cells, co-culturing the applied hepatocytes with the feeder cells, and forming one or more hepatocyte clusters by the co-cultured hepatocytes on the feeder cells, which are attached to the surface. The plated hepatocytes may be used for various purposes, including the preparation of a hepatitis B virus (HBV) infected hepatocyte culture model and drug testing.