СПОСОБ ИНГИБИРОВАНИЯ РАДИАЦИОННО-ИНДУЦИРОВАННОГО УВЕЛИЧЕНИЯ КОЛИЧЕСТВА СТВОЛОВЫХ КЛЕТОК РАКА МОЛОЧНОЙ ЖЕЛЕЗЫ ЧЕЛОВЕКА

Изобретение относится к области медицины, в частности к онкологии, и может быть использовано для ингибирования радиационно-индуцированного увеличения количества CD44+СD24-/low опухолевых стволовых клеток (ОСК). ОСК рака молочной железы человека инкубируют 24 часа до облучения и 48 часов после с водорастворимыми димерными бисбензимидазолами серии DBPA(n) с 1 или 4 метиленовыми группами в составе линкера DBPA, после чего регистрируют снижение радиационно-индуцированного увеличения количества CD44+CD24-/lowОСК, которые являются более радио- и химиорезистентными по сравнению с остальными опухолевыми клетками, а также снижение радиационно-индуцированной экспрессии виментина, который является маркером эпителиально-мезенхимального перехода, обеспечивающего формирование пула ОСК после облучения. Способ позволяет активировать в клетках злокачественных новообразований ряд сигнальных путей и может стать новым инструментом в борьбе со злокачественными новообразованиями. 3 ил., 3 пр.

Способ оценки способности клеток рака молочной железы к дедифференцировке методом образования сфероидов

Изобретение относится к биотехнологии, а именно к способам оценки способности клеток рака молочной железы к дедифференцировке путем культивирования клеток рака молочной железы в полной питательной среде с добавлением эмбриональной телячьей сыворотки, глутамина, антибиотика в стандартных условиях, снятия клеток, дезагрегации и получения одноклеточной суспензии, отбора дифференцированных клеток методом иммунного сортинга, переноса клеток в 6-луночные планшеты с низкой адгезией к пластику, индукции дедифференцировки клеток за счет добавления в среду интерлейкина-6. Изобретение может быть использовано для изучения стволовой пластичности дифференцированных клеток рака молочной железы при исследовании фундаментальных механизмов канцерогенеза, предсказания рисков развития метастазов после оперативного удаления опухоли и для оценки эффективности новых противораковых препаратов, а именно антиметастатических препаратов. 8 ил.

Prostate stem cell

METHOD FOR ISOLATING A CHEMOTHERAPEUTIC AGENT RESISTANT CANCER CELL WITH STEM CELL PROPERTIES

Method for isolating a chemotherapeutic agent resistant cancer cell with stem cell properties

Method for isolating a chemotherapeutic agent resistant cancer cell with stem cell properties

Method for isolating a chemotherapeutic agent resistant cancer cell with stem cell properties

Methods and compositions for treating chronic inflammatory injury, metaplasia, dysplasia and cancers of epithelial tissues

The present disclosure provides methods and formulations for treating a patient suffering from one or more of chronic inflammatory injury, metaplasia, dysplasia or cancer of an epithelial tissue, which method comprises administering to the patient an agent that selectively kills or inhibits the proliferation or differentiation of pathogenic epithelial stem cells (PESCs) relative to normal epithelial stem cells in the tissue in which the PESCs are found. Representative epithelial tissues include pulmonary, genitourinary, gastrointestinal/esophageal, pancreatic and hepatic tissues.

Methods and materials for hematoendothelial differentiation of human pluripotent stem cells under defined conditions

Methods and compositions for differentiating pluripotent stem cells into cells of endothelial and hematopoietic lineages are disclosed.

Method for culturing a subpopulation of circulating epithelial tumour cells from a body fluid

The invention relates to a method for culturing a subpopulation of circulating epithelial tumour cells from a body fluid of a human or animal suffering from an epithelial tumour, wherein cells contained in the body fluid each containing at least one cell nucleus are separated from the body fluid and cultured over at least 24 hours in suspension, with formation of spheroids.

BRAIN TUMOR STEM CELLS

Tumor stem cells have been identified in brain tumors and characterized as expressing the cell surface marker CD133. These stem cells can be employed in prognostic tests for brain tumor patients and in screening tests for potential anti-cancer chemotherapeutics.

METHOD FOR DETECTING OR SEPARATING/OBTAINING CIRCULATING TUMOR CELL EMPLOYING CELL PROLIFERATION METHOD

The present invention addresses the problem of providing a method for detecting or separating/obtaining a circulating tumor cell (CTC), whereby it becomes possible to reliably and steadily detect or separate/obtain a CTC and a circulating tumor stem cell (CTSC) which are present in trace amounts in a biological circulating body fluid, such as blood and lymph fluid, even under the condition where the type of the tumor cell is not identified yet and the tumor cell is present in a trace amount in the biological circulating body fluid. The problem can be solved by a method for detecting or separating/obtaining a CTC and/or a CTSC in a biological circulating body fluid, said method comprising the following treatment steps (1) to (4): (1) a first step of pretreating a sample collected from the biological circulating body fluid to obtain a mononuclear cell phase; (2) a second step of providing a well plate that is prepared by injecting a liquid culture medium comprising a serum-free culture medium ...

DELIVERY OF PAYLOADS TO STEM CELLS

The present disclosure relates to a method of targeting stems cells, in particular non-apoptotic stem cells, employing a GLA domain, capable of binding surface exposed phosphatidyl serine.

COMPOSITIONS ENRICHED IN NEOPLASTIC STEM CELLS AND METHODS COMPRISING SAME

A neoplastic stem cell population enriched for expression of the OCT4 tra nscription factor as well as methods for their identification, isolation and enrichment are described. The OCT4-enriched neoplastic stem cell population is further utilized for the induction and analysis of cancer in an animal. In addition, methods of preventing, abrogating, or inhibiting cancer, tumor growth, and metastasis via OCT4 inhibition are further provided.

NOVEL MEDULLOBLASTOMA-FORMING CELL LINE

Isolated medulloblastoma-forming clonogenic cells are provided which are useful to form stable cell lines as well as a non-human animal models of medulloblastoma th at mimic human medulloblastoma, thereby providing a means to screen for candidate therapeutic compounds.

INHIBITION OF BREAST CARCINOMA STEM CELL GROWTH AND METASTASIS

Disclosed is a method for inhibiting the growth of breast carcinoma stem cells, that express High Molecular Weight-Melanoma Associated Antigen (HMW- MAA). The method comprises administering to an individual a composition comprising an antibody reactive with HMW-MAA or a fragment of such an antibody in an amount effective to inhibit the growth of the breast carcinoma cells. Also provided are methods for inhibiting metastasis of breast carcinomas and methods for identifying HMW-MAA+ breast cancer stem cells.

CANCER STEM CELLS AND USES THEREOF

Disclosed are enriched preparations of neuroblastoma tumor initiating cells (NB TICs). The NB TICs are capable of self-renewal, initiating neuroblastoma tumor growth in vivo and are capable of being passaged in high frequency. These NB TICs have chromosomal abnormalities and are capable of giving rise to secondary tumor spheres. Methods are also disclosed for preparing the enriched preparations of NB TICs, such as from neuroblastoma tumor tissue and metastasized bone marrow. Also disclosed are methods of screening candidate substances to identify therapeutic agents for the treatment of neuroblastoma. Methods are also provided for screening a sample for neuroblastoma, as well as for screening a sample to identify the stage of neuroblastoma present. Kits are also provided for selecting appropriate anti-neuroblastoma compounds for a patient, and utilize isolated compositions of the patients' neuroblastoma tumor initiating cells. In this manner, a customized medicinal profile for the patient ...

COMPOSITIONS AND METHODS FOR TREATING AND DIAGNOSING CANCER

The present invention relates to compositions and methods for characterizing, treating and diagnosing cancer. In particular, the present invention provides a cancer stem cell profile, as well as novel stem cell cancer markers useful for the diagnosis, characterization, prognosis and treatment of cancer and in particular the targeting of solid tumor stem cells.

METHODS AND MEANS RELATED TO CANCER STEM CELLS

This invention relates to the methods for the identification and isolation of cancer stem cells from cultured cancer cell lines. Cell line-derived cancer stem cells isolated using the present methods may be useful, for example, in assays to screen compounds for anti-cancer stem cell activity and in target discovery methods for identifying novel expressed genes and druggable targets. The invention also relates to the screening of compounds for activity against cell line-derived cancer stem cells.

METHODS FOR IDENTIFYING AND TARGETING TUMOR STEM CELLS BASED ON NUCLEAR MORPHOLOGY

Described herein are methods for inhibiting tumor growth comprising targeting a tumor stem cell in the patient with an agent or treatment that chemically modifies a tumor stem cell-specific molecule, thereby preventing proliferation of tumor stem cells.

ISOLATION OF NON-EMBRYONIC STEM CELLS AND USES THEREOF

The invention described herein relates to methods of isolating non-embryonic stem cell, e.g., adult stem cell, from a non-embryonic tissue, e.g., an adult tissue or organ. Non- embryonic stem cells (e.g., adult stem cells) thus isolated from the various tissues or organs can self-renew or propagate indefinitely in vitro, are multipotent and can differentiate into the various differentiated cell types normally found within the tissue or organ from which the stem cells are isolated. In addition, the isolated stem cells can be propagated through clonal expansion of a single isolated stem cell, to produce a clone of which at least about 40%, 70%, or 90% or more cells within the clone can be further passaged as single cell originated clones.

BINDING PROTEINS SPECIFIC FOR LOX1 AND USES THEREOF

This disclosure provides LOX1 (LOX1) binding proteins such as anti-LOX1 antibodies, and compositions and methods for making these binding proteins. In certain aspects the LOX1-binding proteins provided herein, inhibit, or antagonize LOX1 activity. In addition, the disclosure provides compositions and methods for diagnosing and treating conditions associated with atherosclerosis, thrombosis, coronary artery disease (CAD), ischemia (e.g., myocardial ischemia), infarction (e.g., myocardial infarction), acute coronary syndrome (ACS), stroke, reperfusion injury, restenosis, peripheral vascular disease, hypertension, heart failure, inflammation (e.g., chronic inflammation), angiogenesis, preeclampsia, cancer and other LOX1-mediated diseases and conditions.

CANCER STEM CELL ANTIGEN VACCINES AND METHODS

Method of stimulating an immune response (e.g., to treat cancer) include administering to a patient a composition including dendritic cells that present cancer stem cell antigens. Compositions including cancer stem cell antigens are also provided herein.

METHOD FOR THE ISOLATION FOR MAMMALIAN STEM CELLS AND USES THEREOF

The present invention concerns the field of stem cell biology, and in particular relates to a method for producing an isolated bona fide population of mammalian stem cells, and uses of the stem cells thus produced. Human glioblastomas (hGBMs) have now been shown to contain a minor subset of cells bearing the defining features of somatic stem cells (SCs) and the ability to establish, expand and perpetuate these tumors. They are defined stem-like tumor propagating cells (TPCs). This has caused a paradigmatic shift in the way we interpret hGBM physiology, for it identifies TPCs as a major culprit to be tackled for the development of novel therapeutics. It also suggests that studying the regulatory mechanisms of normal neurogenesis may point to specific inhibitors of TPCs.

CHIMERA COMPRISING BACTERIAL CYTOTOXIN AND METHODS OF USING THE SAME

This invention provides, a recombinant polypeptide encoding a chimera. The chimera includes a DNase I fragment or a homologue thereof and a Cdt fragment or a homologue thereof. Further, the invention provides methods, utilizing the recombinant polypeptide encoding the chimera, such as a method for inhibiting the proliferation of a neoplastic cell, a method for treating a neoplastic disease in a human subject, a method for inhibiting or suppressing a neoplastic disease in a human subject, and a method for reducing the symptoms associated with a neoplastic disease in a human subject.

METHODS TO IDENTIFY AND ENRICH FOR POPULATIONS OF OVARIAN CANCER STEM CELLS AND SOMATIC OVARIAN STEM CELLS AND USES THEREOF

The present invention relates to compositions and methods for treating, characterizing and diagnosing ovarian cancer. In particular, the present invention provides methods for treating and/or preventing ovarian cancer in a subject by administering to the subject an effective amount of Mullerian Inhibiting substance and/or an effective amount of an agent that inhibits BCRP1. The present invention further provides methods to identify and/or enrich for populations of ovarian cancer stem cells and populations of somatic ovarian stem cells, in particular, enrichment for populations of coelomic somatic ovarian stem cells, subcoelomic/stromal somatic ovarian stem cells and periphilar medullary somatic ovarian stem cells. The present invention also provides somatic ovarian stem cell markers and ovarian cancer stem cell markers, as well as methods to identify agents which selectively inhibit the proliferation of ovarian cancer stem cells as compared to somatic ovarian stem cells.

PLURIPOTENT CANCER STEM CELL LINE, AND A PRODUCTION METHOD THEREFOR

The present invention relates to a pluripotent cancer stem cell line which is derived from breast-cancer tissue and which expresses breast-cancer stem-cell marking factor. The present invention also relates to a production method for a pluripotent cancer stem cell line, comprising: (1) a stage involving the isolation of breast-cancer cells from previously extracted breast-cancer tissue; (2) a stage involving a first culturing of the isolated breast-cancer cells in a suspended state in a medium for suspension culturing; (3) a stage involving the recovery of the cells in the suspended state from the first culture; and (4) a stage involving the production of a pluripotent cancer stem cell line by passaging the recovered cells for more than a predetermined number of times in a suspended state in a medium for suspension culturing.

Human foreskin fibroblast conditioned media for culturing ES cells

A cell culture comprising human foreskin cells, the human foreskin cells being capable of maintaining stem cells in an undifferentiated state when co-cultured therewith.

Isolation and use of solid tumor stem cells

A small percentage of cells within an established solid tumor have the properties of stem cells. These solid tumor stem cells give rise both to more tumor stem cells and to the majority of cells in the tumor that have lost the capacity for extensive proliferation and the ability to give rise to new tumors. Thus, solid tumor heterogeneity reflects the presence of tumor cell progeny arising from a solid tumor stem cell. This discovery is the basis for solid tumor stem cell compositions, methods for distinguishing functionally different populations of tumor cells, methods for using these tumor cell populations for studying the effects of therapeutic agents on tumor growth, and methods for identifying and testing novel anti-cancer therapies directed to solid tumor stem cells. We have developed a xenograft model in which we have been able to establish tumors from primary tumors via injection of tumors in the mammary gland of severely immunodeficient mice. Xenograft tumors have been established ...

Stem cell with suppressed SOCS and improved immunosuppressive ability and use thereof

The present invention relates to a stem cell having an immunosuppressive ability in which the expression or activity of a suppressor of cytokine signaling (SOCS) is inhibited, and a pharmaceutical composition for inhibiting immunity, which includes the stem cell. In addition, the present invention relates to a composition for inducing the immunosuppressive activity of a stem cell, including a suppressor of cytokine signaling (SOCS) expression or activity inhibitor. The inhibition of suppressor of cytokine signaling (SOCS) expression or activity, according to the present invention, may enhance the immunosuppressive ability of a stem cell, and the stem cell with enhanced immunosuppressive ability may be used as an effective cell therapeutic agent in an autoimmune disease, rejection upon organ transplantation, or an allergic disease.

Composition for inducing dedifferentiation into cancer stem cells comprising ribosome-activating inhibitor as active ingredient, cancer organoid culture method and anticancer drug screening platform based thereon

A composition for inducing dedifferentiation from cancer cells to cancer stem cells comprising a ribosome-activating inhibitor as an active ingredient, a method of culturing a cancer organoid based thereon and an anticancer drug screening platform, and the increase of colorectal cancer stem cell group induced by the exposure of ribosome-inactivating stress was regulated by the ATF3 gene.

Нumаn fоrеskin fibrоblаst соnditiоnеd mеdiа fоr сulturing ЕS сеlls

А сеll сulturе соmprising humаn fоrеskin сеlls, thе humаn fоrеskin сеlls bеing саpаblе оf mаintаining stеm сеlls in аn undiffеrеntiаtеd stаtе whеn со-сulturеd thеrеwith.

METHODS FOR IDENTIFYING, PURIFYING AND ENRICHING IMMATURE OR STEM CANCER-INITIATING CELLS FROM TUMORS AND USE THEREOF

СПОСОБ ОБНАРУЖЕНИЯ ИЛИ ВЫДЕЛЕНИЯ/ПОЛУЧЕНИЯ ЦИРКУЛИРУЮЩЕЙ ОПУХОЛЕВОЙ КЛЕТКИ, ИСПОЛЬЗУЮЩИЙ МЕТОД ПРОЛИФЕРАЦИИ КЛЕТОК

Изобретение относится к медицине, а именно к способу обнаружения циркулирующей опухолевой клетки и/или циркулирующей опухолевой стволовой клетки из биологической циркулирующей жидкости организма. Способ включает этап предварительной обработки образца биологической циркулирующей жидкости организма для получения фазы мононуклеарных клеток, в котором предварительная обработка образца биологической циркулирующей жидкости организма с удалением жидкого компонента и неклеточного компонента, содержащегося в биологической циркулирующей жидкости организма; этап получения луночного планшета, в который введена культуральная среда, содержащая бессывороточную ростовую среду для циркулирующей опухолевой клетки и/или циркулирующей опухолевой стволовой клетки, и посева в него мононуклеарных клеток, полученных на первом этапе, с последующей инкубацией; этап удаления культуральной среды из лунки планшета, полученного путем инкубации на втором этапе; а также этап обнаружения адгезивной опухолевой клетки, прикрепленной ...

СПОСОБЫ ИДЕНТИФИКАЦИИ И ТАРГЕТИНГА РАКОВЫХ СТВОЛОВЫХ КЛЕТОК НА ОСНОВЕ МОРФОЛОГИИ ЯДРА

... 1. Способ ингибирования раковых стволовых клеток, включающий воздействие на раковые стволовые клетки агентом или обработкой, химически модифицирующей специфичную к раковым стволовым клеткам молекулу, тем самым предотвращая пролиферацию раковых стволовых клеток. ! 2. Способ по п.1, в котором специфичная к раковым стволовым клеткам молекула синтезируется и подвергается сегрегации в дочерние колоколообразные ядра. ! 3. Способ по п.1, в котором специфичная к раковым стволовым клеткам молекула представляет собой одноцепочечную ДНК (оцЦНК). ! 4. Способ по п.1, в котором агентом является химический агент. ! 5. Способ по п.1, в котором агентом является фермент. ! 6. Способ по п.1, в котором обработка представляет собой облучение. ! 7. Способ ингибирования опухолевого роста у пациента, включающий воздействие на раковые стволовые клетки у пациента агентом или обработкой, химически модифицирующей специфичную к раковым стволовым клеткам молекулу, тем самым предотвращая пролиферацию раковых стволовых ...

СВЯЗЫВАЮЩИЕ БЕЛКИ, СПЕЦИФИЧНЫЕ В ОТНОШЕНИИ LOX1, И ПУТИ ИХ ПРИМЕНЕНИЯ

КОМПОЗИЦИИ И СПОСОБЫ ПОДАВЛЕНИЯ РАКОВЫХ СТВОЛОВЫХ КЛЕТОК

Prostate cancer stem cells

Culture medium

Novel uses

The present invention relates to the use of a compound represented by the general formula (I), and pharmaceutically acceptable salts thereof, wherein : n is an integer from 2 to 5; p is an integer from 0 to 4; R1 is selected from the group consisting of a hydroxyl group, an alkoxy group, a thiol group, a thioether group, and an amino group, wherein at least two of R1 is a hydroxyl group, a thiol group or an amino group, with the proviso that wherein n is 2, R1 is not on the carbon-2 position and the carbon-5 position of the phenyl group; R2 and R3 are independently a methylene group or a nucleophile, with the proviso that at least one of R2 and R3 is a nucleophile group; Z is an oxygen (O) atom or sulfur (S) atom; T is hydrogen or an optionally substituted aliphatic group; in the manufacture of a medicament for treating a patient in need of cell therapy.

Novel uses

Culture medium

CANCER PROSTATA MAIN CELL

Identification, isolation and elimination of cancer stem cells

Compositions and methods for treating and diagnosing cancer

Methods for identifying, purifying and enriching immature or stem cancer-initiating cells from tumors and use thereof

Methods for identifying, purifying and enriching immature or stem cancer-initiating cells from tumors and use thereof

... 42 Abstract of the invention: The present invention is related to methods for the preparation of cell compositions, isolated compositions obtainable therefrom, related isolated cell compositions, kits and use thereof. More specifically, the present invention provides a method for identifying, purifying and enriching immature or stem cancer-initiating cells in a 5 sample. The cell compositions, related methods and uses according to the present invention are useful in the treatment of cancers and/or the detection of enriching immature or stem cancer-initiating cells, notably cancers of the central and peripheral nervous system, metastasis to the brain.

Compositions and methods for treating and diagnosing cancer

COMPOSITIONS AND METHODS FOR TREATING AND DIAGNOSING CANCER Abstract The present invention relates to compositions and methods for treating, characterizing, and diagnosing cancer. In particular, the present invention provides gene expression profiles associated with solid tumor stem cells, as well as novel stem cell cancer markers useful for the diagnosis, characterization, and treatment of solid tumor stem cells.

Method for detecting or separating/obtaining circulating tumor cell employing cell proliferation method

The present invention addresses the problem of providing a method for detecting or separating/obtaining a circulating tumor cell (CTC), whereby it becomes possible to reliably and steadily detect or separate/obtain a CTC and a circulating tumor stem cell (CTSC) which are present in trace amounts in a biological circulating body fluid, such as blood and lymph fluid, even under the condition where the type of the tumor cell is not identified yet and the tumor cell is present in a trace amount in the biological circulating body fluid. The problem can be solved by a method for detecting or separating/obtaining a CTC and/or a CTSC in a biological circulating body fluid, said method comprising the following treatment steps (1) to (4): (1) a first step of pretreating a sample collected from the biological circulating body fluid to obtain a mononuclear cell phase; (2) a second step of providing a well plate that is prepared by injecting a liquid culture medium comprising a serum-free culture medium ...

Compositions and methods for treating and diagnosing cancer

METHODS AND COMPOSITIONS FOR TREATING CHRONIC INFLAMMATORY INJURY, METAPLASIA, DYSPLASIA AND CANCERS OF EPITHELIAL TISSUES

The present disclosure provides methods and formulations for treating a patient suffering from one or more of chronic inflammatory injury, metaplasia, dysplasia or cancer of an epithelial tissue, which method comprises administering to the patient an agent that selectively kills or inhibits the proliferation or differentiation of pathogenic epithelial stem cells (PESCs) relative to normal epithelial stem cells in the tissue in which the PESCs are found. Representative epithelial tissues include pulmonary, genitourinary, gastrointestinal/esophageal, pancreatic and hepatic tissues.

METHODS FOR PRODUCING HEPATOCYTES

Methods for producing hepatocytes from pluripotent human stem cells are disclosed herein. The stem cells are plated on a cell culture substrate comprising two laminins. The stem cells are then exposed to different cell culture mediums to induce differentiation. The resulting hepatocytes have higher metabolic capacity compared to hepatocytes cultured on different substrates.

METHODS AND PANELS OF COMPOUNDS FOR CHARACTERIZATION OF GLIOBLASTOMA MULTIFORME TUMORS AND CANCER STEM CELLS THEREOF

A method of characterizing a glioblastoma multiforme (GBM) stem cell (GSC), comprising culturing the GSC to provide a culture, contacting a first set of aliquots of the culture with individual compounds selected from a panel of compounds, identifying two or more of the selected compounds that cause more than a threshold level of cell death in the first set of aliquots, and characterizing the GSC as suitable for treatment with one or more combinations comprising the two or more identified compounds. A panel of chemical compounds, the compounds selected by a method comprising surgically resecting the tumor, culturing a GSC derived from GBM tissue derived from a GBM tumor, contacting aliquots thereof with individual compounds selected from a panel of compounds, and identifying two or more of the selected compounds that cause more than a threshold level of cell death in the aliquots, thereby identifying the compounds.

HUMAN CANCER STEM CELLS

This invention discloses isolated populations of human cancer stem cells. Methods for characterizing, isolating and culturing human cancer stem cells are also disclosed. Uses for human cancer stem cells are provided.

INDIVIDUALIZED HIGH PURITY HEPATOCELLULAR CARCINOMA STEM CELLS, METHODS AND USE OF THE SAME

The disclosure provides cancer stem cells, for use in stimulating immune response against a cancer, such as hepatocellular carcinoma (HCC). Methods for preparing and purifying the cancer stem cells are provided.

RAPID METHOD PRODUCTION HIGH PURITY CANCER STEM CELLS AND POPULATION OF HIGH PURITY CANCER STEM CELLS

The disclosure provides reagents, including cells, and related methods, useful for administering to subjects with a neoplastic disorder. The reagents and methods encompass cancer stem cells of enhanced purity. Neoplastic disorder encompasses melanoma, ovarian cancer, colorectal cancer, breast cancer, and lung cancer.

СПОСОБЫ КУЛЬТИВИРОВАНИЯ СТВОЛОВЫХ КЛЕТОК И КЛЕТОК-ПРЕДШЕСТВЕННИКОВ

В изобретении описан способ культивирования, развития или выращивания стволовых или стволовоподобных клеток или индуцированных плюрипотентных стволовых клеток на поверхности, включающий прикрепление клеток к поверхности при помощи лиганда, который связывается с поверхностью и клетками.

СПОСОБЫ ИДЕНТИФИКАЦИИ, ОЧИСТКИ И ОБОГАЩЕНИЯ НЕЗРЕЛЫХ ИЛИ СТВОЛОВЫХ КЛЕТОК-ИНИЦИАТОРОВ ЗЛОКАЧЕСТВЕННОГО РОСТА ИЗ ОПУХОЛЕЙ И ИХ ПРИМЕНЕНИЕ

Настоящее изобретение относится к способам получения композиций клеток, композиций изолированных клеток, получаемых из указанных родственных композиций изолированных клеток, наборам и их применению. Более конкретно, настоящее исследование обеспечивает способ идентификации, очистки и обогащения незрелых или стволовых клеток-инициаторов злокачественного роста в образце. Композиции клеток, относящиеся к ним способы и варианты применения согласно настоящему изобретению могут быть использованы при лечении рака и/или обнаружении увеличивающегося количества незрелых или стволовых клеток-инициаторов злокачественного роста, в частности злокачественных опухолей центральной и периферической нервной системы, метастазов в мозг.

METHOD OF ISOLATION OF CANCER STEM CELLS

POLYPEPTIDE PROBE FOR DETECTING CD133

The present invention relates to a polypeptide probe for detecting CD133 functioning as a neuroglioma stem cell marker. The use of the polypeptide probe of the present invention enables the effective detection of neuroglioma stem cells. To this end, the polypeptide probe includes: (a) a polypeptide backbone made of an amino acid sequence represented by general formula 1, HxGC(GK)yC; and (b) a plurality of monopeptide probes coupled to the backbone and made of a first sequence on the sequence list. COPYRIGHT KIPO 2016 (AA) C (cysteine) : Formation of a probe structure in a cyclic form through a disulfide bond (BB) Peptide^* : CD133 binding peptide sequence, K M P K E N P S S W L S GCGK (FITC) ...

método de isolamento de uma célula de câncer resistente a agente quimioterápico com propriedades de célula tronco

CANCER STEM CELL ANTIGEN VACCINES AND METHODS

Method of stimulating an immune response (e.g., to treat cancer) include administering to a patient a composition including dendritic cells that present cancer stem cell antigens. Compositions including cancer stem cell antigens are also provided herein.

CHIMERA COMPRISING BACTERIAL CYTOTOXIN AND METHODS OF USING THE SAME

This invention provides, a recombinant polypeptide encoding a chimera. The chimera includes a DNase I fragment or a homologue thereof and a Cdt fragment or a homologue thereof. Further, the invention provides methods, utilizing the recombinant polypeptide encoding the chimera, such as a method for inhibiting the proliferation of a neoplastic cell, a method for treating a neoplastic disease in a human subject, a method for inhibiting or suppressing a neoplastic disease in a human subject, and a method for reducing the symptoms associated with a neoplastic disease in a human subject.

METHODS FOR ISOLATING AND USING PITUITARY ADENOMA STEM CELLS AND PITUITARY ADENOMA CELLS

The present invention describes pituitary adenoma stem cells, pituitary carcinoma stem cells, a method of obtaining the stem cells, and a method of using the stem cells. Uses of the pituitary stem cells include but are not limited to producing pituitary hormones and identifying drugs to treat pituitary disease conditions or pituitary-related disease conditions.

Methods of treating pre-malignant ductal cancer with autophagy inhibitors

Described herein are progenitor cancer cells and cell lines isolated from human breast ductal carcinoma in situ (DCIS) lesions and the uses of these cells or cell lines in drug design, drug screening, and monitoring in vivo therapy. The DCIS malignant precursor cells or cell lines are epithelial in origin, are positive for markers of autophagy, show at least one genetic difference from normal cells of said fragment, form 3-D tube-like structures or ball aggregates, or are inhibited in formation of 3-D structures and migration by treatment with chloroquine. In one embodiment, there is a loss of heterozygosity (LOH) that is narrowly confined to a region of chromosome 6p (6p21.1-6p12.3) that contains the SUPT3H gene.

COMPOSITIONS AND METHODS FOR TREATING AND DIAGNOSING CANCER

The present invention relates to compositions and methods for treating, characterizing, and diagnosing cancer. In particular, the present invention provides gene expression profiles and signatures associated with solid tumor stem cells, as well as novel stem cell cancer markers useful for the diagnosis, characterization, prognosis and treatment of solid tumor stem cells. More particularly, the present invention identifies two profiles of cancer stem cells useful for the diagnosis, characterization, and treatment of cancer and cancer metastases. The invention also provides a variety of reagents such as stem cell gene signatures for use in the diagnosis and management of cancer.

METHOD FOR MAINTAINING AND AMPLIFYING COLON CANCER STEM CELLS AND METHOD FOR INDUCING COLON CANCER ORGANOID

The present invention provides a method for maintenance and expansion of a colon cancer stem cell or induction of a colon cancer organoid. In addition, the present invention provides a medicament screening system using a colon cancer stem cell maintained and expanded or a colon cancer organoid induced by the method. 1. A method for maintaining and expanding a colon cancer stem cell or inducing a colon cancer organoid comprising culturing the colon cancer stem cell in the presence of a calcineurin inhibitor.2. The method according to claim 1 , wherein the starting colon cancer stem cell is induced by culturing under conditions where embryonic stem (ES) cells cannot be maintained a colon cancer cell having an exogeneous reprogramming factor introduced thereinto.3. The method according to claim 2 , wherein the starting colon cancer stem cell has a medicament eliminating ability in the presence of an ABC transporter inhibitor at a concentration effective for suppressing a medicament eliminating ability of a colon cancer cell without introduction of an exogeneous reprogramming factor.4. The method according to comprising a step of adhesion-culturing the colon cancer stem cell.5. The method according to comprising a step of three-dimensional culturing the colon cancer stem cell.6. The method according to claim 5 , wherein the three-dimensional culture step is performed after the adhesion-culturing step claim 5 , and one or both of said steps is/are performed in the presence of a calcineurin inhibitor.7. The method according to claim 6 , wherein at least the adhesion culture step is performed in the presence of a calcineurin inhibitor.8. The method according to claim 5 , wherein the three-dimensional culture step is performed by coculturing a mesenchymal stem cell and a vascular endothelial cell.9. The method according to claim 1 , wherein the calcineurin inhibitor is added for a period of 5-25 days.10. The method according to wherein the calcineurin inhibitor is FK506.11. ...

Нumаn fоrеskin fibrоblаst соnditiоnеd mеdiа fоr сulturing ЕS сеlls

А сеll сulturе соmprising humаn fоrеskin сеlls, thе humаn fоrеskin сеlls bеing саpаblе оf mаintаining stеm сеlls in аn undiffеrеntiаtеd stаtе whеn со-сulturеd thеrеwith.

Нumаn fоrеskin fibrоblаst соnditiоnеd mеdiа fоr сulturing ЕS сеlls

А сеll сulturе соmprising humаn fоrеskin сеlls, thе humаn fоrеskin сеlls bеing саpаblе оf mаintаining stеm сеlls in аn undiffеrеntiаtеd stаtе whеn со-сulturеd thеrеwith.

Нumаn fоrеskin fibrоblаst соnditiоnеd mеdiа fоr сulturing ЕS сеlls

А сеll сulturе соmprising humаn fоrеskin сеlls, thе humаn fоrеskin сеlls bеing саpаblе оf mаintаining stеm сеlls in аn undiffеrеntiаtеd stаtе whеn со-сulturеd thеrеwith.

Compositions and methods for treating and diagnosing cancer

The present invention relates to compositions and methods for treating, characterizing, and diagnosing cancer. In particular, the present invention provides gene expression profiles associated with solid tumor stem cells, as well as novel stem cell cancer markers useful for the diagnosis, characterization, and treatment of solid tumor stem cells.

Method for the purification and amplification of tumoral stem cells

The invention concerns a method for the purification and amplification in the undifferentiated state of tumoral stem cells from solid tumours which are most resistant to conventional therapies, aiming at devising new tumour markers and therapeutic targets both for early diagnosis and for targeted therapeutic strategies.

СПОСОБ ДЛЯ ВЫДЕЛЕНИЯ РАКОВЫХ СТВОЛОВЫХ КЛЕТОК

СПОСОБ АДРЕСНОГО ВОЗДЕЙСТВИЯ НА ЭКЗОСОМЫ

СПОСОБЫ ПОЛУЧЕНИЯ ЦИТОКАПСУЛ И ЦИТОКАПСУЛЯРНЫХ ТРУБОК

3D-tissue culture based method to assess mitochondrial impairment

Methods for producing cancer stem cell spheroids

Compositions and methods for treating and diagnosing cancer

HUMAN FORESKIN CELLS SUITABLE FOR CULTURING STEM CELLS

Human foreskin cells suitable for culturing stem cells

Non-static suspension culture of cell aggregates

The invention is directed to compositions of cell aggregates and methods for making and using the cell aggregates where the aggregates comprise cells that are not embryonic stem cells but can differentiate into cell types of at least two of ectodermal, endodermal, and mesodermal embryonic germ layers, e.g., stem cells.

Methods for producing hepatocytes

Methods for producing hepatocytes from pluripotent human stem cells are disclosed herein. The stem cells are plated on a cell culture substrate comprising two laminins. The stem cells are then exposed to different cell culture mediums to induce differentiation. The resulting hepatocytes have higher metabolic capacity compared to hepatocytes cultured on different substrates.

Adhesive signature-based methods for the isolation of cancer-associated cells and cells derived therefrom

The present invention provides methods of isolating a cancer-associated cell, such as a cancer stem cell or tumor initiating cell or a cell derived therefrom, from a mixture of cells, for example, a mixture of adherent cells in culture. Cell isolation is achieved by the application of selective detachment forces.

Methods for culturing stem and progenitor cells

The present application describes a method of culturing, expanding or growing stem or stem-like cells or induced pluripotent stem cells on a surface, including attaching the cells to the surface through a ligand that binds to the surface and the cells.

METHODS FOR PRODUCING CANCER STEM CELL SPHEROIDS

The invention provides a method for producing a population of ready-to-use spheroid forming cancer cells, comprising:(i) growing cancer cells in suspension culture in a first culture medium on one or more first low-adhesion tissue culture plates thereby forming cancer cell spheroids enriched in cancer stem cells; (ii) disaggregating said cancer cell spheroids to form a suspension of single cells enriched in cancer stem cells; (iii) plating said suspension of single cells in a second culture medium on one or more second low-adhesion tissue culture plates; and (iv) freezing said suspension of single cells in said one or more second tissue culture plates, thereby producing a population of ready-to-use spheroid forming cancer cells. Also provided are cell populations produced by the method and kits for growing cancer cell spheroids, including for use in screening of test compound.

METHOD FOR ISOLATING AND DETECTING CANCER STEM CELLS

La présente invention concerne l'utilisation in vitrod'au moins une lectine pour le marquage de cellules souches cancéreuses d'organes cibles de cancers hormono-dépendants, choisie parmiles lectines Maackia amurensislectin II (MAH-II), Euonymus europaeuslectin (EEL), Psophocarpus tetragonolobuslectin I (PTL-I) et Griffonia Simplicifolialectin II (GSL-II), pour obtenir des cellules souches cancéreuses d'organes cibles de cancers hormono- dépendants marquées, dans un échantillon biologique. notamment au moins deux lectines choisies parmi MAH-II, EEL, PTL-I et GSL-II, en particulier les deux lectines MAH-II et EEL.

METHOD FOR ISOLATING CANCER STEM CELLS

La présente invention concerne l'utilisation d'une lectine reconnaissant le motif fucosea 1-2 galactose en tant que premier moyen de marquage et éventuellement un second moyen de marquage des cellules souches cancéreuses colorectales, notamment une lectine reconnaissant l'antigène T pour la mise en uvre d'un procédé de détection et éventuellement d'isolement des cellules souches cancéreuses colorectales, un procédé de de détection et éventuellement d'isolement des cellules souches cancéreuses colorectales à des fins de recherche et une méthode de diagnostic in vitro du risque de récidive du cancer colorectal et/ou de l'agressivité du cancer colorectal pour définir une valeur pronostique pour l'adaptation thérapeutique d'un cancer colorectal, ainsi qu'un kit comprenant une lectine reconnaissant le motif fucose a 1-2 galactose et une lectine reconnaissant l'antigène T.

PROSPECTIVE IDENTIFICATION AND CHARACTERIZATION OF BREAST CANCER STEM CELLS

Human breast tumors contain heterogeneous cancer cells. Using an animal xenograft model in which human breast cancer cells were grown in immunocompromised mice, we found that only a small minority of breast cancer cells had the capacity to form new tumors. The ability to form new tumors was not a stochastic property, rather certain populations of cancer cells were depleted for the ability to form new tumors, while other populations were enriched for the ability to form new tumors. Tumorigenic cells could be distinguished from non-tumorigenic cancer cells based on surface marker expression. We prospectively identified and isolated the tumorigenic cells as CD44+CD24-/loLINEAGE-. As few as 100 cells from this population were able to form tumors the animal xenograft model, while tens of thousands of cells from non-tumorigenic populations failed to form tumors. The tumorigenic cells could be serially passaged, each time generating new tumors containing an expanded numbers of CD44+CD24-/lowLineage-tumorigenic ...

TUMOR-INITIATING CELLS AND METHODS FOR USING SAME

Isolated and enriched tumor-initiating cell popula-tions, methods for preparing the same, and uses thereof.

PROSTATE STEM CELLS AND USES THEREOF

Prostate stem cells and prostate cancer stem cells and their use in treating prostate cancer and regenerating prostate tissue are disclosed.

METHOD OF ISOLATION OF CANCER STEM CELLS

La présente invention concerne l'utilisation d'une lectine reconnaissant le motif fucose α 1-2 galactose en tant que premier moyen de marquage et éventuellement un second moyen de marquage des cellules souches cancéreuses colorectales, notamment une lectine reconnaissant l'antigène T pour la mise en œuvre d'un procédé de détection et éventuellement d'isolement des cellules souches cancéreuses colorectales, un procédé de de détection et éventuellement d'isolement des cellules souches cancéreuses colorectales à des fins de recherche et une méthode de diagnostic in vitro du risque de récidive du cancer colorectal et/ou de l'agressivité du cancer colorectal pour définir une valeur pronostique pour l'adaptation thérapeutique d'un cancer colorectal, ainsi qu'un kit comprenant une lectine reconnaissant le motif fucose α 1-2 galactose et une lectine reconnaissant l'antigène T.

ANIMAL MODEL FOR AMPLIFYING HUMAN OR ANIMAL CIRCULATING TUMOR CELLS

Methods for culturing stem and progenitor cells

줄기 세포 및 전구 세포를 배양하는 방법

... 본원은 표면 및 세포에 결합하는 리간드를 통해 세포를 표면에 부착시킴을 포함하여, 표면에서 줄기 또는 줄기-유사 세포 또는 유도된 다능성 줄기 세포를 배양하거나, 확장시키거나 성장시키는 방법을 기술한다.

INDUCED MALIGNANT STEM CELLS

METHOD FOR CULTURING A SUBPOPULATION OF CIRCULATING EPITHELIAL TUMOUR CELLS FROM A BODY FLUID

Proteínas de unión específicas para lox1 y sus usos

...

Esta descripción propociona proteínas de unión a LOX1 (LOX1) tales como anticuerpos contra LOX1 y composiciones y métodos para producir estas proteínas de unión. En ciertos aspectos, las proteínas de unión a LOX1 proporcionadas en la presente inhiben la actividad de LOX1 o son antagonistas respecto a ella. Además, la descripción proporciona composiciones y métodos para diagnosticar y tratar afecciones asociadas con las aterosclerosis, trombosis, arteriopatía coronaria (CAD), isquemia (por ejemplo, isquemia de miocardio), infarto (por ejemplo, infarto de miocardio), síndrome coronario agudo (ACS), accidente cerebrovascular, lesión por reperfusión, reestenosis, enfermedad vascular periférica, hipertensión, insuficiencia cardíaca, inflamación (por jemplo, inflamación crónica), angiogénesis, preenclampsia, cáncer y otras enfermedades mediadas por LOX1.

Method for the Purification and Amplification of Tumoral Stem Cells

The invention concerns a method for the purification and amplification in the undifferentiated state of tumoral stem cells form solid tumours which are most resistant to conventional therapies, aiming at devising new tumour markers and therapeutic targets both for early diagnosis and for targeted therapeutic strategies.

Glioblastoma multiforme-reactive antibodies and methods of use thereof

The present disclosure is generally related to antibodies that bind specifically to glioblastoma multiforme (GBM) cells. In particular, the present disclosure provides compositions comprising human single chain or full-length antibodies that bind tumor cells. Additionally the present disclosure provides methods of using the anti-GBM antibodies.

Use of catalytic antioxidant to preserve stem cell phenotype and control cell differentiation

Methods are disclosed herein for maintaining stem cells in an undifferentiated state in vitro. The methods include contacting the stem cells with an effective amount of a catalytic antioxidant. Also disclosed are methods for the increasing the number of stem cells in vitro while maintaining the stem cells in an undifferentiated state. The methods include contacting the stem cells with an effective amount of a catalytic antioxidant and an effective amount of one or more growth factors that promotes the expansion of the stem cells.

Methods of isolating stem cells

The present inventors discovered for the first time that labeling cell nuclei makes it possible to efficiently isolate stem cells. Namely, it was elucidated that stem cells with labeled nuclei remained labeled even after cell division, and showed self-renewing and long-living abilities characteristic of stem cells. Efficient isolation of stem cells is possible, for instance, by labeling the nuclear of each cell in a heterogeneous cellular group followed by selecting those cells that maintain a labeled state even after cell division. The present invention provides methods for enabling visualization of stem cells of animal tissues in a living state by labeling using the essential functions of the stem cells, and methods for simply and easily isolating the stem cells in a fresh state without using at all genetic manipulation or artificial markers.

In vitro assays for enriching and determining the clonogenic potential of stem cells

A method of enriching stem or progenitor cells that includes growing a heterogeneous cell sample comprising stem and/or progenitor cells on a first substrate that is hydrophobic and has an elastic modulus less than about 100 MPa; recovering the heterogeneous cell sample from the first substrate; growing the recovered heterogeneous cell sample on a second substrate that is hydrophilic and has an elastic modulus higher than the elastic modulus of the first substrate to produce a subpopulation of nonadherent cells and a subpopulation of adherent cells; and recovering the nonadherent cell subpopulation, which is enriched for stem and/or progenitor cells. The invention also relates to a method of determining the clonogenic potential of a cell, such as a cancer stem cell enriched using the enrichment method described herein.

Identification and Enrichment of Cell Subpopulations

Markers useful for the identification, characterization and, optionally, the enrichment or isolation of tumorigenic cells or cell subpopulations are disclosed.

SCREENING METHODS, COMPOSITIONS IDENTIFIED THEREBY, TOOLS USEFUL FOR THE IDENTIFICATION THEREOF, AND CELL POPULATIONS PRODUCED THEREBY

In accordance with one aspect of the present invention, methods have been developed for identifying compositions which support the culture of defined cell populations. In accordance with another aspect of the present invention, methods have been developed for identifying compositions which promote differentiation of defined cell populations. In accordance with yet another aspect of the present invention, methods have been developed for identifying compositions which induce apoptosis of defined cell populations. In accordance with still another aspect of the present invention, methods have been developed for identifying compositions which promote cell senescence of defined cell populations. In accordance with still another aspect of the present invention, methods have been developed for identifying media which modulate the retardation of cell growth of defined cell subpopulation(s). In accordance with further aspects of the present invention, there are provided novel compositions identified by invention methods. Also provided are various uses of the novel compositions identified by invention methods, and novel cell populations produced employing same. In accordance with still another aspect of the present invention, methods have been developed for identifying compositions which support the culture of aberrant cell populations. In accordance with yet another aspect of the present invention, methods have been developed for identifying compositions which promote differentiation of aberrant cell populations. In accordance with still another aspect of the present invention, methods have been developed for identifying compositions which induce apoptosis of aberrant cell populations. 2. The method of claim 1 , wherein one or more of the plurality of components which comprise each complex microenvironment closely resemble component(s) found in the in vivo environment in which said cell subpopulation(s) normally reside(s) claim 1 , or in the in vivo environment of a species ...

INDUCED MALIGNANT STEM CELLS OR PRE-INDUCTION CANCER STEM CELLS CAPABLE OF SELFREPLICATION OUTSIDE OF AN ORGANISM, PRODUCTION METHOD FOR SAME, AND PRACTICAL APPLICATION FOR SAME

The present invention provides an induced cancer cell capable of self-replication in vitro which is useful in cancer therapy research and the research for cancer-related drug discovery, processes for production thereof, cancer cells induced by the malignant cells, and applications of these cells. 1. An induced cancer stem cell which is an induced precancer stem cell or an induced malignant stem cell , wherein the induced cancer stem cell has the following two characteristics:(1) expressing the six genes POU5F1, NANOG, SOX2, ZFP42, LIN28, and TERT; and(2) having an aberration which is either (a) a mutation in an endogenous tumor suppressor gene or (b) increased expression of an endogenous cancer-related gene.2. The induced cancer stem cell according to claim 1 , wherein the self-renewal related genes as referred to in (1) above are expressed in the induced cancer stem cell in amounts ranging from one-eighth to eight times the amounts of the genes that are expressed in an embryonic stem cell.3. The induced cancer stem cell according to or claim 1 , which is an induced precancer stem cell.4. The induced cancer stem cell according to claim 3 , wherein the tumor suppressor gene referred to (a) is APC or RB1.5. The induced cancer stem cell according to or claim 3 , which is an induced malignant stem cell.6. The induced cancer stem cell according to claim 5 , wherein the cancer-related gene referred to (b) is within at least one group of genes selected from the groups of genes consisting of a group of genes related to angiogenesis claim 5 , a group of cancer-related pathway genes claim 5 , a group of genes related to stromal barrier claim 5 , a group of genes related to epithelial-mesenchymal transition claim 5 , a group of genes related to stomach cancer claim 5 , a group of genes related to autonomous growth claim 5 , a group of genes related to TGF β/BMP signaling claim 5 , a group of genes related to tissue invasion/metastasis claim 5 , a group of genes related to Wnt ...

METHODS OF USING CDK8 ANTAGONISTS

Provided herein are CDK8 antagonists and methods of using the same. 1: A method of screening for and/or identifying a CDK8 antagonist which promotes cell differentiation said method comprising: contacting a reference cell , wherein the reference cell is a stem cell and/or a cancer stem cell , with a CDK8 candidate antagonist , wherein the CDK8 candidate antagonist binds CDK8 , and whereby differentiation of the reference cell into a differentiated cell identifies the CDK8 candidate antagonist as a CDK8 antagonist which promotes cell differentiation.2: The method of claim 1 , wherein the reference cell is a cancer stem cell.3: The method of claim 2 , wherein the differentiated cell is a goblet cell and/or enterocyte cell.4: The method of claim 1 , wherein the CDK8 candidate antagonist is an antibody claim 1 , binding polypeptide claim 1 , small molecule claim 1 , or polynucleotide.5: A method of inducing differentiation comprising contacting the cell with an effective amount of CDK8 antagonist.6: The method of claim 5 , wherein the cell is a stem cell.7: The method of claim 5 , wherein the cell is a cancer stem cell.8: A method of treating a cancer cell claim 5 , wherein the cancer cell differentially expresses one or more biomarkers of a CDK8 gene signature (e.g. claim 5 , compared to a reference sample claim 5 , reference cell claim 5 , reference tissue claim 5 , control sample claim 5 , control cell claim 5 , control tissue claim 5 , or internal control (e.g. claim 5 , housekeeping gene)) claim 5 , the method comprising providing an effective amount of a CDK8 antagonist.919-. (canceled)20: The method of claim 1 , wherein differential expression of one or more biomarkers of the CDK8 gene signature is elevated expression of one or more CDK8-induced biomarkers of the CDK8 gene signature and/or reduced expression of one or more CDK8-repressed biomarkers of the CDK8 gene signature.21: (canceled)22: The method of claim 8 , wherein the one or more biomarkers of the CDK8 ...

METHOD FOR ISOLATING A CHEMOTHERAPEUTIC AGENT RESISTANT CANCER CELL WITH STEM CELL PROPERTIES

The invention relates to the use of encapsulates of cancer cells, in agarose coated, agarose containing beads, for isolating chemotherapeutic resistant cells which have at least one stem cell property, such as expression of OCT4. The cells thus isolated are also a feature of the invention, as is a method for screening for potential therapeutic 1. A method for isolating a chemotherapeutic agent resistant cancer cell with stem cell properties , comprising:(i) encapsulating a sample of cancer cells in an agarose coated, agarose containing bead;(ii) contacting said bead with at least one anti-cancer chemotherapeutic agent for a time sufficient to kill at least a portion of said cancer cells;(iii) removing any surviving cells from said bead;(iv) assaying said surviving cell for at least one stem cell property, and(v) isolating any cells which exhibit said stem cell property from any cells which do not.2. The method of claim 1 , wherein said stem cell property is expression of OCT4.3. The method of claim 1 , wherein said chemotherapeutic agent is Docetaxel.4. The method of claim 1 , wherein said cancer cells are mammalian cells.5. The method of claim 4 , wherein said mammalian cells are human cells.6. An isolated claim 4 , chemotherapeutic resistant cancer cell which exhibits at least one stem cell property.7. The isolated chemotherapeutic resistant cancer cell of claim 6 , wherein said stem cell property is expression of OCT4.8. The isolated chemotherapeutic resistance cancer cell of claim 6 , wherein said cancer cell is a mammalian cell.9. The isolated chemotherapeutic resistant cancer cell of claim 8 , wherein said mammalian cell is a human cell.10. The isolated chemotherapeutic resistant cancer cell of claim 5 , wherein said cell is resistant to Docetaxel.11. A method for determining if a substance of interest has anti-cancer efficacy claim 5 , comprising (i) contacting said substance to a sample of cancer cells encapsulated in an agarose containing claim 5 , agarose ...

TUMOR STEM CELLS

Tumor stem cells can be obtained by culturing a tumor cell population, and exposing the cultured tumor cell population to free radicals. In certain embodiments, the free radical agent can be a nitric oxide (NO) donor. In one embodiment, the free radical agent can be Diethylenetriamine NONOate (DETA NONOate) or agents that constitutively increase cellular nitric oxide, such as phosphodiesterase inhibitors or L-arginine, or agents that increase NO synthase in the population. The methods can further include inducing stem cells present in the population to expand and/or inducing dedifferentiation of tumor cells into tumor stem cells. Additionally, the present invention provides methods of selecting stem cells from a tumor cell population. Another aspect provides methods of screening for anti-tumor stem cell teherapeutic compounds by providing high nitric oxide (HNO) tumor cells, exposing the HNO cells to at least one compound, assessing one or more indicators of HNO cell health and determining toxicity of the compound to HNO tumor cells. 1. A method of obtaining tumor stem cells comprisingculturing a cell population, andexposing the cultured cell population to free radicals to obtain the tumor stem cells.2. The method of wherein the step of exposing the cultured cell population to free radicals comprises exposing the cultured cell population to nitrogen-based free radicals.3. The method of wherein nitrogen-based free radicals includes nitric oxide (NO).4. The method of wherein nitrogen-based free radicals includes a NO donor.5. The method of wherein the step of exposing the cultured cell population further comprises exposing the population to Diethylenetriamine NONOate (DETA NONOate).6. The method of wherein the step of exposing the cultured cell population further comprises increasing NO synthase in the population. (Original) The method of wherein the step of exposing the cultured cell population to free radicals comprises exposing the cultured cell population to ...

Glioma Stem Cells and Methods for Obtaining Them

The present invention relates to a glioma stem cell population, wherein the glioma stem cells do not present a telomerase activity and to a method for obtaining such cells. 1. An isolated glioma stem cell (GSC) population , wherein the glioma stem cells of the population do not present a telomerase activity.2. The glioma stem cell population according to claim 1 , wherein the glioma stem cells of the population display heterogeneous telomere lengths and/or an intracellular colocalization of telomeric DNA and promyelocytic leukemia (PML) nuclear bodies.3. The glioma stem cell population according to claim 1 , wherein the population generates glioblastoma tumors upon administration to an animal.4. The glioma stem cell population according to claim 1 , wherein less than 1% of the glioma stem cells of the population express CD133.5. The glioma stem cell population according to claim 1 , wherein from 30% to 70% of the glioma stem cells of the population express CD15.6. The glioma stem cell population according to claim 1 , wherein more than 90% of the glioma stem cells of the population express Sox2.7. The glioma stem cell population according to claim 1 , deposited at the Collection Nationale de Culture de Micro-organismes (CNCM) claim 1 , Institut Pasteur claim 1 , Paris claim 1 , France claim 1 , under the Budapest Treaty claim 1 , on Jul. 30 claim 1 , 2010 claim 1 , under accession number CNCM I-4348 claim 1 , or on Jun. 21 claim 1 , 2011 claim 1 , under accession number CNCM I-4496.8. A method of obtaining a glioma stem cell population claim 1 , comprising:culturing cells dissociated from a glioma tumor sample obtained from an individual, in a neural stem cell culture medium comprising epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF-2) to obtain neurosphere populations;determining that the tumor sample or the cells dissociated from the tumor sample do not present a telomerase activity and/or selecting a neurosphere population which do not present a ...

CANCER STEM CELL MASS AND PROCESS FOR PRODUCTION THEREOF

The purpose of the present invention is to provide: a cancer stem cell mass from which cells incapable of forming cancer are substantially removed and which has a characteristic property of reproducing a layered structure of a cancer tissue; a process for producing the cancer stem cell mass; and use of the cancer stem cell mass. For achieving the purpose, the present inventors grew a human cancer tissue repeatedly in a NOG mouse, separated cancer cells from the grown cancer tissue, and made a comparison of various cancer cell culture processes with each other. As a result, a cancer stem cell composition which is homogeneous and is substantially free of the coexistence of cells capable of forming cancer and cells incapable of forming cancer in a mixed state can be produced successively by employing an attached culture process using a serum-free stem cell culture medium rather than a generally employed floating culture process, and consequently the present invention has been accomplished. 1. A population of cancer stem cells from which cells with no cancer-forming ability have been substantially removed , wherein the population is characterized by reproducing the hierarchical structure of a cancer tissue.2. The cancer stem cell population of claim 1 , wherein the cancer stem cells are derived from a human tumor tissue.34-. (canceled)5. The cancer stem cell population claim 1 , which is substantially homogeneous.6. The cancer stem cell population of claim 1 , wherein the frequency of cancer stem cells in extreme limiting dilution analysis is 1/20 or higher.7. The cancer stem cell population of claim 1 , which comprises 1×10or more cancer stem cells.8. The cancer stem cell population of claim 1 , which is produced by a method comprising the step of adherently culturing a cell group containing cancer stem cells.9. The cancer stem cell population of claim 1 , which is produced by a method comprising the steps of:(1) transplanting a cell group containing cancer stem cells ...

Spontaneously immortalized prostate cancer cell line

This disclosure provides prostate cancer cell lines established from spontaneously immortalized, extremely tumorigenic and clonogenic primary prostate tumor. These cell lines represent unique cancer cell and cancer stem cell (CSC) models for preclinical prostate cancer studies and CSC-targeted drug development, which is of high value for pharmaceutic companies producing anti-cancer agents, as well as for the broad range of basic and translational research focused on cancer cell and CSC biology, stem cell behavior, cancer development and metastasis.

Non-Static Suspension Culture of Cell Aggregates

The invention is directed to compositions of cell aggregates and methods for making and using the cell aggregates where the aggregates comprise cells that are not embryonic stem cells but can differentiate into cell types of at least two of ectodermal, endodermal, and mesodermal embryonic germ layers, e.g., stem cells.

COMPOSITIONS AND METHODS FOR TREATING AND DIAGNOSING CANCER

The present invention relates to compositions and methods for characterizing, treating and diagnosing cancer. In particular, the present invention provides a cancer stem cell profile, as well as novel stem cell cancer markers useful for the diagnosis, characterization, prognosis and treatment of cancer and in particular the targeting of solid tumor stem cells. 168.-. (canceled)69. A method for obtaining from a head and neck tumor a cellular composition comprising head and neck cancer stem cells and non-tumorigenic head and neck tumor cells , wherein at least 75% of the tumor cells in the composition are tumorigenic head and neck cancer stem cells and 25% or less of the tumor cells in the composition are non-tumorigenic head and neck tumor cells , the method comprising:(a) obtaining a dissociated mixture of head and neck tumor cells;(b) separating the mixture of head and neck tumor cells into a first fraction comprising at least 75% head and neck cancer stem cells and 25% or less non-tumorigenic head and neck tumor cells and a second fraction of head and neck tumor cells depleted of head and neck cancer stem cells, wherein the separating comprises contacting the mixture with a reagent against CD44; and{'sup': '+', '(c) collecting the first fraction, wherein the head and neck cancer stem cells in the first fraction are CD44.'}70. The method of claim 69 , wherein the separating is performed by flow cytometry claim 69 , fluorescence activated cell sorting (FACS) claim 69 , panning claim 69 , affinity chromatography or magnetic selection.71. The method of claim 69 , wherein the separating is performed by FACS analysis.72. The method of claim 69 , wherein the separating further comprises contacting the mixture with a reagent against ESA; and collecting the first fraction claim 69 , wherein the head and neck cancer stem cells in the first fraction are CD44ESA.73. A method for preparing a population of head and neck tumor cells enriched for head and neck cancer stem cells ...

METHODS FOR CULTURING STEM AND PROGENITOR CELLS

The present application describes a method of culturing, expanding or growing stem or stem-like cells or induced pluripotent stem cells on a surface, including attaching the cells to the surface through a ligand that binds to the surface and the cells. 123-. (canceled)241. A method of harvesting cells from cells grown according to the method of claim , comprising adding a competing molecule that binds to the ligand so that the cells are released from binding to the ligand or the surface.251. A method of harvesting cells from cells grown according to the method of claim , comprising cleaving a linker bound to the surface that is directly or indirectly attached to the cells , so that the cells are released from the surface.26. A method of identifying state of differentiation of cells comprising using anti-MUC1* antibody to bind to the cells , wherein positive signal for anti-MUC1* antibody indicates puripotent cell state , and cells showing binding to non-clipped MUC1 indicates differentiated cell state.27. The method according to claim 26 , comprising separating cells from a mixed population of stem and stem-like cells or induced pluripotent stem cells and newly differentiating cells claim 26 , comprising using anti-MUC1* antibody to bind to the cells claim 26 , wherein positive signal for anti-MUC1* antibody indicates a pluripotent cell state claim 26 , and cells showing binding to non-clipped MUC1 indicates differentiated cell state.28. The method according to claim 27 , further comprising contacting the cells with antibodies to stem or stem-like cell or induced pluripotent stem cell markers claim 27 , wherein positive signal for a stem or stem-like cell or induced pluripotent stem cell marker indicates the presence of pluripotent stem cell state.29. The method according to claim 28 , wherein the cells are contacted with anti-MUC1* and anti-Tra 1-81 claim 28 , anti-Tra 1-60 claim 28 , SSEA3 or SSEA4 antibodies.30. A method of detecting cancer stem cells using anti- ...

INDUCED PLURIPOTENT STEM CELL MODEL OF CHRONIC MYELOID LEUKEMIA REVEALED OLFACTOMEDIN 4 AS A NOVEL THERAPEUTIC TARGET IN LEUKEMIA STEM CELLS

Disclosed herein are compositions and methods to treat and reduce therapeutic resistance in chronic myelogenous leukemia. Also disclosed herein are methods to generate leukemia stem cell like cells (iLSCs) generated from CML patient-derived iPSCs, and methods for utilizing iLSCs in screens to identify modulators of CML drug resistance and gene targets that underlie CML drug resistance. 1. A kit for treatment of chronic myeloid leukemia (CML) , comprising therapeutically effective doses of a tyrosine kinase inhibitor and an inhibitor of OLFM4.2. The kit of claim 1 , wherein the OLFM4 inhibitor comprises an siRNA against the OLFM4 mRNA claim 1 , a knock-down vector against OLFM4 mRNA claim 1 , or an antibody against OLFM4 protein.3. The kit of claim 1 , wherein the tyrosine kinase inhibitor inhibits BCR-ABL tyrosine kinase.4. The kit of claim 1 , wherein the tyrosine kinase inhibitor is imatinib.5. The kit of claim 1 , wherein the tyrosine kinase inhibitor is selected from the group consisting of imatinib claim 1 , nilotinib claim 1 , dasatinib claim 1 , bosutinib claim 1 , ponatinib claim 1 , bafetinib and saracatinib.6. A method for treating CML comprising administering to a subject in need thereof a therapeutically effective amount of an inhibitor of OLFM4.7. The method of claim 6 , wherein the OLFM4 inhibitor comprises an siRNA against OLFM4 mRNA claim 6 , an RNAi expression vector against OLFM4 mRNA claim 6 , or an antibody against OLFM4 protein.8. The method of claim 6 , further comprising administering a therapeutically effective amount of a tyrosine kinase inhibitor.9. The method of claim 8 , wherein the tyrosine kinase inhibitor inhibits BCR-ABL tyrosine kinase.10. The method of claim 9 , wherein the tyrosine kinase inhibitor is imatinib.11. The method of claim 6 , wherein administration of the OLFM4 inhibitor and the tyrosine kinase inhibitor is concurrent.12. A method of identifying a modulator of CML drug resistance claim 6 , comprising:(i) culturing ...

Bioreactor System

A three dimensional cell culture and bioreactor system is provided. The system comprises one or more cell culture chamber. Each cell culture chamber comprises an inlet port and an outlet port in fluid communication with the cell culture chamber. The cell culture chambers may be segregated or in fluid communication with one another. The systems may be used to conduct drug efficacy test, isolate certain cell types from a complex tissue sample of multiple cell types, allow for the ex vivo culturing of patient tissue samples to help guide the course of treatment, and conduct co-culture experiments.

Cancer stem cells

Cancer stem cell populations characterized by expression of CD44 hi , ABCG2, β-catenin, CD117, CD133, ALDH, VLA-2, CD166, CD201, IGFR, and/or EGF1R, and methods of isolating and using the same.

INDIVIDUALIZED HIGH PURITY HEPATOCELLULAR CARCINOMA STEM CELLS, METHODS AND USE OF THE SAME

The disclosure provides cancer stem cells, for use in stimulating immune response against a cancer, such as hepatocellular carcinoma (HCC). Methods for preparing and purifying the cancer stem cells are provided. 1. An immunogenic composition comprising dendritic cells activated ex vivo by tumor antigens derived from a population of purified hepatocellular carcinoma cancer stem cells (HCC-CSCs).2. The immunogenic composition of claim 1 , wherein the tumor antigens comprise cell extracts of the HCC-CSCs.3. The immunogenic composition of claim 1 , wherein the tumor antigens comprise lysates of the HCC-CSCs.4. The immunogenic composition of claim 1 , wherein the tumor antigens comprise intact HCC-CSCs.5. The immunogenic composition of claim 4 , wherein the intact HCC-CSCs are rendered non-proliferative.6. The immunogenic composition of wherein the intact HCC-CSCs are rendered non-proliferative by irradiation.7. The immunogenic composition of claim 5 , wherein the intact HCC-CSCs are rendered non-proliferative by exposure of the HCC-CSCs to a nuclear cross-linking agent.8. The immunogenic composition of claim 1 , further comprising a pharmaceutically acceptable carrier or excipient.9. The immunogenic composition of claim 1 , further comprising an adjuvant.10. The immunogenic composition of claim 9 , wherein the adjuvant is granulocyte macrophage colony stimulating factor.11. The immunogenic composition of claim 1 , wherein the composition comprises activated dendritic cells and HCC-CSCs.12. The immunogenic composition of claim 1 , wherein the HCC-CSCs are in form of HCC-CSC spheroids.13. The immunogenic composition of claim 1 , wherein the HCC-CSCs are in form of early HCC-CSCs.14. The immunogenic composition of claim 1 , wherein the HCC-CSCs are in form of mixed HCC-CSCs.15. The immunogenic composition of claim 1 , wherein the HCC-CSCs are in form of EMT-HCC-CSCs.16. A method of treating hepatocellular carcinoma in a subject in need thereof claim 1 , comprising ...

CELL AND UTILIZATION THEREOF

Provided is an epithelial tissue stem cell derived from an adult, which lacks at least one tumor suppressor gene. 1. An epithelial tissue stem cell derived from an adult , which lacks at least one tumor suppressor gene.2. The cell according to claim 1 , which is an Epithelial cell adhesion molecule (EpCAM)CD31CD45.3. The cell according to claim 1 , which expresses EpCAM and Stem cell antigen-1 (Sca-1) claim 1 , or expresses Surfactant protein-C (SPC) and Secretoglobin family 1A member 1 (CC10).4. The cell according to claim 1 , which expresses Lymphocyte antigen 6 family member D (Ly6D).5. The cell according to claim 1 , which expresses Cytokeratin 19 (CK19).6. The cell according to claim 1 , wherein the tumor suppressor gene is selected from the group consisting of Ink4a/Arf claim 1 , p53 claim 1 , and PTEN.7. The cell according to claim 1 , which further expresses at least one oncogene.8. The cell according to claim 7 , wherein the oncogene is a mutated KRAS gene claim 7 , a mutated EGFR gene claim 7 , an ALK fusion gene claim 7 , or a ROS1 fusion gene.9. A method for producing a cell culture containing 50% or more of epithelial tissue stem cells derived from an adult claim 1 , the method comprising 3D culturing the cell according to .10. A cell culture produced by the method according to .11. An organoid comprising the cell according to .12. A method for screening a cell transforming agent claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'culturing the cell according to in the presence of a test substance; and'}evaluating characteristics of the cells.13. A method for screening an anticancer agent claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00006', 'claim 6'}, 'culturing the cell according to in the presence of a test substance; and'}measuring proliferation of the cell.14. A method for producing a cancer-bearing non-human animal model claim 7 , comprising transplanting the cell according to into a non-human animal.15. The method for ...

INDIVIDUALIZED HIGH PURITY COLON CARCINOMA STEM CELLS, METHODS AND USE OF THE SAME

The disclosure provides cancer stem cells, for use in stimulating immune response against a cancer, such as colon carcinoma (CC). Methods for preparing and purifying the cancer stem cells are provided. 1. An immunogenic composition comprising dendritic cells activated ex vivo by tumor antigens derived from a population of purified colon carcinoma cancer stem cells (CC-CSCs).2. The immunogenic composition of claim 1 , wherein the tumor antigens comprise cell extracts of the CC-CSCs.3. The immunogenic composition of claim 1 , wherein the tumor antigens comprise lysates of the CC-CSCs.4. The immunogenic composition of claim 1 , wherein the tumor antigens comprise intact CC-CSCs.5. The immunogenic composition of claim 4 , wherein the intact CC-CSCs are rendered non-proliferative.6. The immunogenic composition of wherein the intact CC-CSCs are rendered non-proliferative by irradiation.7. The immunogenic composition of claim 5 , wherein the intact CC-CSCs are rendered non-proliferative by exposure of the cells to a nuclear cross-linking agent.8. The immunogenic composition of claim 1 , further comprising a pharmaceutically acceptable carrier and/or excipient.9. The immunogenic composition of claim 1 , further comprising an adjuvant.10. The immunogenic composition of claim 9 , wherein the adjuvant is granulocyte macrophage colony stimulating factor.11. The immunogenic composition of claim 1 , wherein the composition comprises activated dendritic cells and CC-CSCs.12. The immunogenic composition of claim 1 , wherein the CC-CSCs are in form of CC-CSC spheroids.13. The immunogenic composition of claim 1 , wherein the CC-CSCs are early CC-CSCs.14. The immunogenic composition of claim 1 , wherein the CC-CSCs are mixed CC-CSCs.15. The immunogenic composition of claim 1 , wherein the CC-CSCs are epithelial to mesenchymal transitioned colon carcinoma cancer stem cells (EMT-CC-CSCs).16. A method of treating colon carcinoma in a subject in need thereof claim 1 , comprising ...

PRODUCTION METHOD FOR ARTIFICIAL CANCER STEM CELL AND INDUCED DIFFERENTIATION METHOD THEREFOR

It is intended to provide a cancer stem cell and a method for preparing the same. The present invention provides a method for preparing a pluripotent cancer stem cell, comprising transferring Oct3/4, Sox2, Klf4, and c-Myc genes to an immortalized epithelial cell. The present invention also provides a pluripotent cancer stem cell as prepared by the above method. 124.-. (canceled)25. A method for preparing a cancer stem cell in vitro , comprising transferring Oct3/4 , Sox2 , Klf4 , and c-Myc genes to an immortalized epithelial cell , selecting an alkaline phosphatase staining-positive cell among colonies formed with the cells transduced with the four genes to obtain a pluripotent cancer stem cell , culturing the pluripotent cancer stem cell in a suspension system to form an embryoid body , and culturing the embryoid body in an adhesion system.26. A cancer stem cell prepared by the method of .27. A method for preparing a cancer cell in vitro claim 26 , comprising differentiating the pluripotent cancer stem cell of .28. The method of claim 27 , comprising culturing the cancer stem cell in an adhesion system.29. A cancer cell prepared by the method of .30. A method for screening for a substance having an anticancer effect by using the cell of or .31. A method of claim 25 , wherein the immortalized epithelial cell is selected from the group consisting of human mammary epithelial cells MCF-10A claim 25 , human prostatic epithelial cells RWPE-1 claim 25 , and human skin keratinocytes HaCaT. The present invention relates to a production method for an artificial cancer stem cell and an induced differentiation method therefor.In recent years, stem cell biology research has advanced, and strongly suggested the existence of cancer stem cells in solid tumors or hematological tumors. These cancer stem cells are found as small subsets, also called side populations (SPs), in solid tumors. This population, in order to maintain the tumor mass, gives rise to many cancer cells, while ...

TRANSFORMED HUMAN PLURIPOTENT STEM CELLS AND ASSOCIATED METHODS

The present disclosure provides transformed human pluripotent stem cell (t-hPSC). t-hPSCs are not dependent on Oct4 for renewal and survival, however exhibit a sensitivity to reduced levels of the transcription factor Nanog. Also provided are methods of culturing cells for use in a cell-based screening assay comprising placing one or more transformed human pluripotent stem cells into a receptacle and culturing said stem cells in the receptacle to form a monolayer of stem cells without cell overlap. Methods of screening compounds using t-hPSCs are also described. 1. A transformed human pluripotent stem cell (t-hPSC).2. The transformed human pluripotent stem cell of claim 1 , wherein the cell co-expresses FGFR1 and IGFR1.3. The transformed human pluripotent stem cell of claim 1 , wherein the cell does not require bFGF for maintenance of an undifferentiated state.4. The transformed human pluripotent stem cell of claim 3 , wherein the cell maintains expression of SSEA3 in the absence of bFGF.5. The transformed human pluripotent stem cell of claim 1 , wherein the self-renewal and survival of the cell is independent of Oct4.6. The transformed human pluripotent stem cell of claim 1 , wherein the cells require Nanog for self-renewal and cell survival.7. The transformed human pluripotent stem cell of claim 1 , wherein the cell exhibits reduced neuronal differentiation or reduced hematopoietic potential in vitro compared to a normal human pluripotent stem cell.8. The transformed human pluripotent stem cell of claim 1 , wherein t-hPSC-derived neural precursors do not form metastasis in vivo.9. The transformed human pluripotent stem cell of claim 1 , wherein the cell comprises an amplification of at least 0.8 megabase at 20q11.1-11.2.10. The transformed human pluripotent stem cell of claim 1 , wherein the cell comprises a deletion at 5q34a-5q34b;5q3 and a mosaic gain of chromosome 12.11. The transformed human pluripotent stem cell of claim 1 , wherein the cell can be passaged ...

Cell Cryopreservation Protective Composition, Use Thereof, and Cell Cryopreservation Method

The present application relates to a cell cryopreservation protective composition, the use of the composition, and a cell cryopreservation method. The cell cryopreservation protective composition comprises a zwitterionic molecule having the general formula R—N(CH)—(CH)—R, wherein Ris a linear or branched alkyl having 1 to 10 carbon atoms, and is optionally substituted with a substituent; Ris any negatively charged group selected from the group consisting of —COO, —SO, —SO, and (I); and Ris a group selected from the group consisting of (methyl)acryloyloxyalkyl, alkyl and alkenyl; and the zwitterionic molecule having general formula R—N(CH)—(CH)—Ris preferably a betaine compound. The cell cryopreservation protective composition can carry out cell cryopreservation in a non-toxic and efficient manner, and results in an extremely high post-thaw cell survival rate and does not require stepwise cryopreservation. After cell recovery, the cells can be used directly or after being slightly diluted. 3. The cell cryopreservation protective composition according to claim 2 , wherein said zwitterionic molecules are 420 to 1120 parts by mass with respect to 100 parts by mass of said nutrient ingredient for cells; and{'sub': 1', '3', '2', '2', 'n', '2, 'sup': '+', 'in said structure of general formula R—N(CH)—(CH)—R,'}{'sub': '1', 'said Ris linear or branched alkyl having 1 to 3 carbon atoms, and is optionally substituted with a substituent selected from the group consisting of (meth)acryloylamino, (meth)acryloyloxy, hydroxyl, hydroxyalkyl having 1 to 5 carbon atoms, and alkoxyl having 1 to 5 carbon atoms,'}{'sub': 3', '2', '2', 'n, 'the hydrogen atoms in said (CH)and (CH)may each independently be optionally substituted with a substituent selected from the group consisting of alkyl having 1 to 5 carbon atoms, hydroxyl, hydroxyalkyl having 1 to 5 carbon atoms, alkoxyl having 1 to 5 carbon atoms, and a polyalkylene oxide group, and'}n is an integer from 1 to 3.5. The cell ...

CANCER STEM CELL

The purpose of the present invention is to provide: a cancer stem cell having an excellent ability to form tumor tissue; and a method for establishing the cancer stem cell. Cells having a low metabolic function of 26S proteasome are isolated and concentrated from a cancer stem cell group containing cancer stem cells, thereby obtaining cancer stem cells which can form tumor tissue in a living body even when the number of cells is 200 or less. 1. A cancer stem cell , which is capable of forming tumor tissue with 200 or less cells in vivo.2. The cancer stem cell according to claim 1 , which is capable of forming tumor tissue with one cell in vivo.3. The cancer stem cell according to claim 1 , which highly expresses CD44v9.41. The cancer stem cell according to claim 1 , which is Panc- 3-4 CST 001 line (Accession number: NITE BP-02449).5. A method for separating a cancer stem cell claim 1 , comprising a step of separating and enriching a cell having a low metabolic function of 26s proteosome from a cancer cell group containing the cancer stem cell.6. The method for separating a cancer stem cell according to claim 5 , wherein the step of separating and enriching a cell having a low metabolic function of 26s proteosome is conducted by transducing a labeled ornithine decarboxylase-degron into the cancer cell group.7. The method for separating a cancer stem cell according to claim 5 , comprising further separating a cell highly expressing CD44v9 from the cell having a low metabolic function of 26s proteosome claim 5 , the cell being enriched.8. A method for screening a cancer therapeutic agent claim 1 , comprising screening a cancer therapeutic agent using the cancer stem cell according to .9. A method for evaluating efficacy of a cancer therapeutic agent claim 1 , comprising evaluating efficacy of a cancer therapeutic agent using the cancer stem cell according to .10. The cancer stem cell according to claim 2 , which highly expresses CD44v9.11. The method for separating a ...

METHOD FOR ISOLATING AND DETECTING CANCER STEM CELLS

Disclosed is the in vitro use of at least one lectin for marking cancer stem cells of hormone-dependent cancer target organs, selected from the lectins lectin II (MAH-II), lectin (EEL), lectin I (PTL-I) and lectin II (GSL-II), in particular at least two lectins selected from MAH-II, EEL, PTL-I and GSL-II, in particular the two lectins MAH-II and EEL, in order to obtain cancer stem cells of labeled hormone-dependent cancer target organs in a biological sample. 113-. (canceled)14Maackia amurensisEuonymus europaeusPsophocarpus tetragonolobusGrijfonia simplicifolia. An in vitro method of labeling by using of at least one lectin for the labeling of cancer stem cells of hormone-dependent cancer target organs , chosen from the lectins lectin II (MAH-II) , lectin (EEL) , lectin I (PTL-I) and lectin II (GSL-II) , to obtain labeled cancer stem cells of hormone-dependent cancer target organs , in a biological sample.15. The in vitro method according to claim 14 , wherein at least two lectins are chosen from MAH-II claim 14 , EEL claim 14 , PTL-I and GSL-II.16. The in vitro method according to claim 15 , wherein at least two lectins are used claim 15 , said at least two lectins being in equal amount or in unequal amount claim 15 , in particular in unequal amount in a weight ratio of 2:1 claim 15 , and preferably both lectins being MAH-II/EEL in unequal amount in a 2:1 weight ratio.17. The in vitro method according to claim 14 , wherein the MAH-II lectin recognizes O-linked glycans claim 14 , the PTL-I lectin recognizes-linked glycans claim 14 , the EEL lectin recognizes galactosylated glycans claim 14 , and the GSL-II lectin recognizes N-linked glycans.18. The in vitro method according to claim 17 , wherein the MAH-II lectin recognizes the disialyl-T group [NeuAc α2-3Gal α1-3 (NeuAc α2-6) GalNAc].19. The in vitro method according to claim 17 , wherein the PTL-I lectin recognizes the Gal α 1-3 (Fuc α 1-2) Gal and GalNAc α 1-3 (Fuc α 1-2) Gal groups of the B and A antigens.20. ...

Macrophage Enrichment of Cancer Stem Cells

Disclosed are culture systems and methods that can enrich a cancer cell population in cancer stem cells. The culture system can be utilized for study of cancer stem cells in general as well as for cancer drug screening. The methods and systems include co-culturing of a cancer cell population and programable macrophages, which can lead to proliferation of the cancer stem cells of the population and depletion of differentiated cancer cells of the population. The methods and systems can provide a more reliable and accurate model for preclinical drug toxicity testing. 1. A method for culturing a population of cancer cells , the method comprising:encapsulating a cancer cell population in a biocompatible matrix, the cancer cell population including cancer stem cells, wherein at least a portion of the biocompatible matrix is non-adherent to the cancer stem cells and the differentiated cancer cells, and wherein the biocompatible matrix is inert to the cancer stem cells and to the differentiated cancer cells;encapsulating macrophages in the biocompatible matrix, the cancer cell population and the macrophages being in chemical communication with one another; andculturing the cancer cell population and the macrophages while held in chemical communication with one another in the biocompatible matrix, wherein over the course of the culturing, the cancer cell population is enriched in the cancer stem cells and depleted in the differentiated cancer cells.2. The method of claim 1 , the differentiated cancer cells comprising breast claim 1 , lung claim 1 , colon claim 1 , gastric claim 1 , liver claim 1 , thyroid claim 1 , bladder claim 1 , oral claim 1 , ovarian claim 1 , or nasal cancer cells.3. The method of claim 1 , further comprising deriving the cancer cell population from a subject.4. The method of claim 1 , further comprising obtaining the cancer cell population from a cancer cell line.5. The method of claim 1 , the method comprising culturing the cancer cell population and ...

Methods and compositions for targeting cancer stem cells

Aspects of the disclosure relate to methods that involve activating the Protein Kinase A (PKA) pathway to induce cancer stem cells (CSCs) to undergo a mesenchymal to epithelial transition. Methods provided herein are useful, in some embodiments, because they render CSCs amenable to treatment with conventional cancer therapies. In some embodiments, methods are provided that involve assaying PKA pathway activity to identify compounds that selectively target CSCs.

Method for detecting or separating/obtaining circulating tumor cell employing cell proliferation method

A method for detecting or separating/obtaining CTC, which is capable of reliably and stably detecting or separating/obtaining a circulating tumor cell and a circulating tumor stem cell present in trace amounts in a biological circulating fluid such as blood or lymph, even in the state where the cancer type of the tumor cells cannot be determined yet and the state where the tumor cells are present in trace amounts in the biological circulating fluid. The method is attained by detecting or separating/obtaining CTC and/or CTSC in a biological circulating fluid, comprising treatment steps (1) to (4): (1) pretreating a sample from the biological circulating fluid to obtain a mononuclear cell phase; (2) providing a well plate in which a culture medium consisting of a serum-free cell growth medium for circulating tumor cell and/or circulating tumor stem cell has been injected, and seeding thereto the mononuclear cells obtained in step (1), followed by incubation; (3) removing the culture medium from a well of the plate obtained by the incubation in step (2); and (4) detecting or separating/obtaining an adherent tumor cell attached to the well of the plate after step (3).

METHODS AND PRODUCTS FOR TRANSFECTION

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed. 19.-. (canceled)10. A method for reprogramming a differentiated cell to a less differentiated state , comprising:(a) providing a differentiated cell;(b) culturing the differentiated cell; and wherein the one or more synthetic RNA molecules include at least one RNA molecule encoding one or more reprogramming factors selected from the group consisting of Oct4 protein, Sox2 protein, Klf4 protein, c-Myc protein, I-Myc protein, Tert protein, Nanog protein, and Lin28 protein;', 'wherein the transfecting results in the cell expressing the one or more reprogramming factors to result in the cell being reprogrammed to a less differentiated state; and', 'wherein step (c) occurs in the presence of a medium containing ingredients that support reprogramming of the differentiated cell to a less differentiated state., '(c) transfecting the differentiated cell with one or more synthetic RNA molecules,'}11. The method of claim 10 , wherein the differentiated cell is derived from a biopsy.12. The method of claim 11 , wherein the differentiated cell is derived from a dermal punch biopsy sample.13. The method of claim 10 , wherein the differentiated cell is from a human subject.14. The method of claim 10 , wherein the differentiated cell is a skin cell.15. The method of claim 10 , further comprising contacting the cell with at least one member of the group: poly-L-lysine claim 10 , poly-L-ornithine claim 10 , RGD peptide claim 10 , fibronectin claim 10 , vitronectin claim 10 , collagen claim 10 , and laminin.16. The method of claim 10 , wherein the one or more synthetic RNA molecules contain at least one of a pseudouridine or a 5-methylcytidine residue.17. The method of claim 10 , wherein ...

High purity ovarian cancer stem cells for active autologous immune therapy

The disclosure provides cancer stem cells, for use in stimulating immune response against a cancer, such as ovarian carcinoma. Methods for preparing and purifying the cancer stem cells are provided.

INDUCED MALIGNANT STEM CELLS

PROBLEM 1. An induced malignant stem cell capable of in vitro proliferation that is characterized by satisfying the following two requirements:(1) having at least one aberration selected from among (a) an aberration of methylation (high or low degree of methylation) in a tumor suppressor gene or a cancer-related genetic region in endogenous genomic DNA, (b) a somatic mutation of a tumor suppressor gene or a somatic mutation of an endogenous cancer-related gene in endogenous genomic DNA, (c) abnormal expression (increased or reduced/lost expression) of an endogenous oncogene or an endogenous tumor suppressor gene, (d) abnormal expression (increased or reduced/lost expression) of a noncoding RNA such as an endogenous cancer-related microRNA, (e) abnormal expression of an endogenous cancer-related protein, (f) an aberration of endogenous cancer-related metabolism (hypermetabolism or hypometabolism), (g) an aberration of endogenous cancer-related sugar chain, (h) an aberration of copy number variations in endogenous genomic DNA, and (i) instability of microsatellites in endogenous genomic DNA in an induced malignant stem cell; and(2) expressing genes including POU5F1 gene, NANOG gene, SOX2 gene, and ZFP42 gene.3. The induced malignant stem cell capable of in vitro proliferation according to claim 1 , wherein the somatic mutation of a tumor suppressor gene or a somatic mutation of an endogenous cancer-related gene in endogenous genomic DNA under (1)(b) above comprises a passenger mutation.4. The induced malignant stem cell capable of in vitro proliferation according to claim 1 , wherein the somatic mutation of a tumor suppressor gene or a somatic mutation of an endogenous cancer-related gene in endogenous genomic DNA under (1)(b) above is a driver mutation.6. The induced malignant stem cell capable of in vitro proliferation according to claim 1 , wherein the abnormal expression (increased or reduced/lost expression) of an endogenous oncogene or an endogenous tumor ...

Composition and kit for separating cancer cell, and method of separating cancer cell by using the composition and kit

Provided is a composition and a kit for separating a cancer stem cell or a circulating tumor cell, a method of separating a cancer stem cell or a circulating tumor cell in a biological sample, and a method for diagnosing metastatic cancer.

Method for treatment of gm1 gangliosidosis

The present invention relates to a method for preparing a GM1 gangliosidosis human cell model based on induced pluripotent stem cells (iPSCs) and iPSCs originated neural progenitor cells, and a use of the GM1 model above for the development of a GM1 gangliosidosis treating agent. The iPSCs originated from GM1 patient fibroblasts can be differentiated into neural progenitor cells (NPCs) and neurosphere cells that can emulate the characteristics shown in GM1 patient, so that the said cells can be efficiently used for the investigation of intracellular GM1 symptoms such as the GM1 gangliosidosis and lysosome accumulation and the gene expression pattern change. So, the GM1 cell model of the present invention can be efficiently used for the study of GM1 development mechanism and the study for the development of a therapeutic agent for the disease. The present inventors also established the inflammasome inhibitor rhIL1RA or Z-YVAD-FMK by using the above GM1 cell model and further confirmed that it can be efficiently used as a relieving/treating agent of GM1 gangliosidosis. In addition, the molecular symptoms of GM1 patient could be reproduced in the transformed cells having the E186A mutation which is newly identified as the GM1 gangliosidosis causing protein mutation. Therefore, the mutant cells containing the induced E186A mutation can be efficiently used as the GM1 gangliosidosis cell model.

BIOREACTOR SYSTEM

A three dimensional cell culture and bioreactor system is provided. The system comprises one or more cell culture chamber. Each cell culture chamber comprises an inlet port and an outlet port in fluid communication with the cell culture chamber. The cell culture chambers may be segregated or in fluid communication with one another. The systems may be used to conduct drug efficacy test, isolate certain cell types from a complex tissue sample of multiple cell types, allow for the ex vivo culturing of patient tissue samples to help guide the course of treatment, and conduct co-culture experiments. 1. A method for testing the efficacy of pharmaceutical agents using three-dimensional cell culturing systems comprising;culturing a target cell population in a cell culture chamber of a three-dimensional (3D) cell culturing system;exposing the target cell population in the cell culture chamber to a pharmaceutical agent for a defined period of time and under a defined set of exposure conditions; anddetermining a cell viability or other measurable parameter for the target cell population, wherein the target cell population is responsive to the pharmaceutical agent if the cell viability or other measurable parameter of the target cell population is less than the cell viability or other measurable parameter of a control cell population;wherein the cell culture chamber of the 3D cell culturing system comprises an inlet port and an outlet port that are in communication with an interior volume of the cell culture chamber, the interior volume of the cell culture chamber housing a cell-scaffold construct and the target cell population.2. The method of claim 1 , wherein the target cell population is a cancer cell population claim 1 , and the pharmaceutical agent is an anti-cancer agent.3. The method of claim 2 , wherein the cancer cell population is obtained from a patient biopsy sample.4. The method of claim 1 , wherein the 3D culture system further comprises two or more cell culture ...

METHOD OF OBTAINING A CELL POPULATION CONTAINING CANCER STEM CELLS

The present invention relates to a method of obtaining a cell population containing cancer stem cells, which comprises culturing iPS cells in the presence of culture supernatant of cancer cells. 1. A method of obtaining a cell population containing cancer stem cells , which comprises culturing iPS cells in 5 the presence of culture supernatant of cancer cells.2. The method of claim 1 , wherein the iPS cells are human cells.3. The method of claim 1 , wherein the period of culture is 2 weeks to 2 months.4. The method of claim 1 , wherein the cell population obtained by the method contains cancer cells.5. A cell population obtained by the method of .6. A method of obtaining cancer stem cells claim 5 , which comprises isolating cancer stem cells contained in the cell population of .7. Cancer stem cells obtained by the method of .8. A method of screening an anti-cancer agent claim 6 , which comprises the steps of:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(1) bringing a cell population containing cancer stem cells obtained by the method of or cancer stem cells obtained by isolating cancer stem cells contained in the cell population containing cancer stem cells into contact with a candidate of an anti-cancer agent, and'}(2) determining the effect of the candidate on the cell population or the cells.9. The method of claim 8 , further comprising the step of selecting a candidate which inhibits proliferation of the cell population or the cells.10. The method of claim 8 , which comprises the step of bringing the cell population containing cancer stem cells into contact with a candidate of an anti-cancer agent.11. A method of concentrating CD44-expressing cells claim 8 , which comprises culturing a cell population containing CD44-expressing cells in the presence of hyaluronic acid under the non-adherent condition.12. The method of claim 11 , wherein the cell population is a human cell population.13. The method of claim 11 , wherein the period of the culture is 2 weeks ...

3-D TISSUE CULTURE BASED METHOD TO ASSESS MITOCHONDRIAL IMPAIRMENT

The present invention relates to a method and/or assay for the assessment of the metabolic effect of a candidate compound. The method and/or assay comprises exposing one or more 3-dimensional cell culture or tissue to one or more candidate compounds, and measuring, in at least one 3-dimensional cell culture or tissue, the effect of such exposure on the 3-dimensional cell culture or tissue-specific respiration rate (MTSRR) (FIG. B). 1. Method and/or assay for the assessment of the metabolic effect of a candidate compound , which method and/or assay comprisesa) exposing one or more 3-dimensional cell culture or tissue to one or more candidate compounds, andb) measuring, in at least one 3-dimensional cell culture or tissue, the effect of such exposure on the 3-dimensional cell culture or tissue-specific respiration rate (MTSRR),c) measuring, in the same or at least one other 3-dimensional cell culture or tissue, the effect of such exposure on cell viability and/or the cytotoxic effect (CV-CYT) in one or more cells of at least one 3-dimensional cell culture or tissue, andd) comparing the effects of steps b) and c) to provide an estimate on the mitochondrial toxicity of the one or more candidate compounds,{'sub': 50', '5O', '_', 'MTSRR', '50', '5O', '_', 'CV/CYT, 'wherein the mitochondrial toxicity is determined on the basis of the quantitative relationship between the ICwith respect to MTSRR (IC) and the ICwith respect to CV-CYT (IC).'}2. (canceled)3. The method and/or assay according to claim 1 , wherein said method and/or assay is used to determine the mitochondrial toxicity of a candidate compound.4. The method and/or assay according to claim 1 , wherein said method and/or assay is used to determine the chemotherapeutic efficacy of a candidate compound.5. The method and/or assay of claim 1 , wherein the 3-dimensional cell culture or tissue is a spheroidal cell culture.6. The method and/or assay of any of claim 1 , wherein the 3-dimensional cell culture or tissue ...

Optimized cancer stem cell vaccines

The present invention provides cancer stem cell vaccines useful for treating or preventing a variety of tumors, as well as related methods of producing cancer stem cells and antigens thereof and producing vaccine adjuvants with enhanced activity for use with the stem cell vaccines.

Three Dimensional Matrix for Cancer Stem Cells

Synthetic inert 3D gel culture systems are described that can be finely tuned to exhibit desired and predetermined physical, chemical, mechanical, and biochemical properties. The culture system can be utilized to study the effect of microenvironmental factors on cancer cell response, and in particular on cancer stem cell (CSC) response. Cancer cells can be encapsulated in a crosslinked gel system having a narrow range of predetermined gel stiffness. One or more biochemical factors including peptides that can affect the growth, development, and/or proliferation of CSCs can be incorporated in the system to examine the effects of the factor(s) on the encapsulated cells with regard to growth, proliferation, size, etc. 1. A three dimensional hydrogel matrix comprising a crosslinked inert synthetic polymer that is absent of ligands that can interact with cell surface receptors , the hydrogel matrix further comprising a peptide conjugated to the matrix , the three dimensional hydrogel matrix having an elastic modulus of from 2 kilopascals to 70 kilopascals.2. The three dimensional hydrogel matrix of claim 1 , wherein the three dimensional hydrogel matrix has an elastic modulus of from about 2.5 kilopascals to about 10 kilopascals.3. The three dimensional hydrogel matrix of claim 1 , the peptide affecting the growth claim 1 , development claim 1 , and/or proliferation of a cancer stem cell.4. The three dimensional hydrogel matrix of claim 1 , wherein the peptide comprises a CD44 binding peptide or a mutant thereof claim 1 , an integrin binding peptide or a mutant thereof claim 1 , or a heparin binding peptide or a mutant thereof.5. The three dimensional hydrogel matrix of claim 1 , wherein the peptide comprises RLVSYNGIIFFLK (SEQ ID NO.: 17) claim 1 , VLFGFLKIYSRIN (SEQ ID NO.: 18) claim 1 , GRGDS (SEQ ID NO.: 19) claim 1 , GRDGS (SEQ ID NO.: 20) claim 1 , WQPPRARI (SEQ ID NO.: 21) claim 1 , or RPQIPWAR (SEQ ID NO.: 22).6. The three dimensional hydrogel matrix of further ...

Culture Medium

The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids.

Method for generating cancer stem cells from immortalized cell lines

A method is described for generating a novel cancer stem cell line that possesses characteristics associated with stem cells, by co-culturing a human immortalized cell line and bone marrow-derived mesenchymal stem cells, and the novel cancer stem cell line established thereby. This method is able to readily generate cancer stem cells that are low in the level of structural chromosomal aberrations and are excellent in oncogenicity, and is effectively applicable to the development of anti-cancer drugs and personalized drugs.

METHOD FOR PRODUCING CANCER STEM CELLS

The present invention relates to a method of producing cancer stem cells that comprises culturing a living cell population containing cancer cells in the presence of a gel substance to obtain a living cell population containing cancer stem cells, wherein the gel substance is a material that induces expression of osteopontin in at least a portion of cells contained in the living cell population. Moreover, the present invention relates to an agent for inducing conversion of cancer cells to cancer stem cells that comprises a gel substance that induces expression of osteopontin in at least a portion of the cells contained in a living cell population. The gel substance is a synthetic polymer gel composed of, for example, double network gel, PNaSS gel, PCDME gel, PA gel, RAMPS gel, PDMA gel or PAAc gel. The present invention provides means and a method that enable the preparation of cancer stem cells in a relatively short period of time and at a relatively low culturing cost without requiring expensive equipment. 1. A method of producing cancer stem cells , comprising:culturing a living cell population containing cancer cells in the presence of a gel substance to obtain a living cell population containing cancer stem cells, whereinthe gel substance is a material that induces expression of osteopontin in at least a portion of cells contained in the living cell population, and by culturing in the presence of the gel substance, at least a portion of cancer cells contained in a living cell population containing cancer cells is converted to cancer stem cells.2. The production method according to claim 1 , wherein the gel substance is a synthetic polymer gel composed of double network gel claim 1 , PNaSS gel claim 1 , PCDME gel claim 1 , PA gel claim 1 , PAMPS gel claim 1 , PDMA gel or PAAc gel.3. The method according to claim 1 , wherein the living cell population containing cancer stem cells contains cells exhibiting a spherical structure.4. The method according to claim 1 , ...

HUMAN TISSUE STEM CELL AND USE THEREOF

A human tissue stem cell in which a gene of interest is introduced into a gene locus of a stem cell marker gene. 1. A human tissue stem cell in which a gene of interest is introduced into a gene locus of a stem cell marker gene.2. The human tissue stem cell according to claim 1 ,wherein the stem cell marker gene is a Leucine-Rich Repeat-Containing G Protein-Coupled Receptor 5 (LGR5) gene.3. The human tissue stem cell according to claim 2 ,wherein the gene of interest is introduced into exon 18 of an Lgr5 gene locus.4. The human tissue stem cell according to claim 1 ,wherein the gene of interest is a reporter gene.5. The human tissue stem cell according to claim 1 ,wherein the gene of interest is a first reporter gene, anda second reporter gene that is operated by a forced expression promoter is further introduced.6. The human tissue stem cell according to claim 1 , which is a stem cell derived from cancer tissue.7. A method of introducing a gene of interest into a gene locus of a stem cell marker gene of a human tissue stem cell claim 1 , the method comprising:introducing the gene of interest into the gene locus of the stem cell marker gene of a cultured organoid containing the human tissue stem cell by genome editing.8. The method according to claim 7 ,wherein introducing the gene of interest is performed by electroporation, andthe method further comprises culturing the cultured organoid at 30° C. for 2 days, after introducing the gene of interest.9. The method according to claim 7 ,wherein the gene locus of the stem cell marker gene is an Lgr5 gene locus.10. The method according to claim 9 ,wherein the gene of interest is introduced into exon 18 of the Lgr5 gene locus.11. The method according to claim 7 ,wherein the gene of interest is a gene encoding a cell lineage marker-inducing protein.12. A method of testing whether or not a human tissue stem cell prepared by the method according to is a stem cell claim 11 , comprising:detecting self-replication and ...

CELL CULTURE MEDIUM, CULTURE METHOD, AND ORGANOID

The present invention provides a cell culture medium with which serum-free long-term culture of an epithelial stem cell, an epithelial cancer cell, or a tissue containing at least one thereof can be achieved. The cell culture medium of the present invention includes: a Wnt agonist composed of a complex of Wnt protein and afamin, which is a substance capable of stabilizing the Wnt protein, and R-spondin; and at least one selected from the group consisting of a mitogenic growth factor, a bone morphogenetic protein (BMP) inhibitor, a transforming growth factor-β (TGF-β) inhibitor, and a p38 inhibitor. 1. A cell culture medium comprising:a Wnt agonist including a complex of a Wnt protein and an afamin, which is a substance capable of stabilizing the Wnt protein; andat least one selected from the group consisting of an R-spondin, a mitogenic growth factor, a bone morphogenetic protein (BMP) inhibitor, a transforming growth factor-β (TGF-β) inhibitor, and a p38 inhibitor.2. The cell culture medium according to claim 1 , wherein the Wnt protein is at least one selected from the group consisting of Wnt1 claim 1 , Wnt2 claim 1 , Wnt2b claim 1 , Wnt3 claim 1 , Wnt3a claim 1 , Wnt4 claim 1 , Wnt5a claim 1 , Wnt5b claim 1 , Wnt6 claim 1 , Wnt7a claim 1 , Wnt7b claim 1 , Wnt8a claim 1 , Wnt8b claim 1 , Wnt9a claim 1 , Wnt9b claim 1 , Wnt10a claim 1 , Wnt10b claim 1 , Wnt11 claim 1 , and Wnt16.3. The cell culture medium according to claim 1 , wherein the R-spondin is at least one selected from the group consisting of R-spondin 1 claim 1 , R-spondin 2 claim 1 , R-spondin 3 claim 1 , and R-spondin 4.4. The cell culture medium according to claim 1 , wherein the mitogenic growth factor is an epidermal growth factor (EGF).5. The cell culture medium according to claim 1 , wherein the BMP inhibitor is Noggin.6. The cell culture medium according to claim 1 , wherein the TGF-β inhibitor is A83-01.7. The cell culture medium according to claim 1 , which is a serum-free medium.8. A method ...

METHODS AND PRODUCTS FOR TRANSFECTION

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed. 19.-. (canceled)10. A method for reprogramming a non-pluripotent cell comprising:(a) providing a non-pluripotent cell;(b) culturing the non-pluripotent cell; and(c) transfecting the non-pluripotent cell with a synthetic RNA molecule, wherein:the synthetic RNA molecule encodes one or more reprogramming factor(s) selected from the group consisting of Oct4 protein, Sox2 protein, Klf4 protein, c-Myc protein, 1-Myc protein, Tert protein, Nanog protein, and Lin28 protein,the transfecting results in the non-pluripotent cell expressing the one or more reprogramming factor(s) which reprograms the non-pluripotent cell; andstep (c) is performed without using irradiated human neonatal fibroblast feeder cells and occurs in the presence of a medium containing ingredients that support reprogramming of the non-pluripotent cell.11. The method of claim 10 , wherein the non-pluripotent cell is derived from a biopsy.12. The method of claim 10 , wherein the non-pluripotent cell is harvested from a human subject.13. The method of claim 11 , wherein the non-pluripotent cell is harvested from a dermal punch biopsy sample.14. The method of claim 10 , wherein the non-pluripotent cell is a skin cell.15. The method of claim 10 , further comprising contacting the cell with at least one member of the group: poly-L-lysine claim 10 , poly-L-ornithine claim 10 , RGD peptide claim 10 , fibronectin claim 10 , vitronectin claim 10 , collagen claim 10 , and laminin.16. The method of claim 10 , wherein the synthetic RNA molecule contains at least one of a pseudouridine or a 5-methylcytidine residue.17. The method of claim 10 , wherein the medium is substantially free of immunosuppressants. The present ...

CELL CULTURE SUBSTRATE, MANUFACTURING METHOD THEREFOR, AND USE THEREOF

The present invention provides a cell culture substrate comprising a polymer formed of a cyclosiloxane compound and a manufacturing method therefor, and a method for preparing a cell spheroid type of cell aggregate or induced pluripotent stem cells using the cell culture substrate. The cell spheroid type of cell aggregate can be easily formed by culturing cells on the cell culture substrate of the present invention, and further, the cell culture substrate can be utilized as a cell culture platform for preparing the induced pluripotent stem cells. 1. A cell culture substrate comprising a polymer formed of a cyclosiloxane compound.3. The cell culture substrate of claim 2 , wherein the cyclosiloxane has (n+1) or (n+2) Calkenyl at the Rpositions.4. The cell culture substrate of claim 1 , wherein the cyclosiloxane compound is selected from the group consisting of{'sub': 1-10', '2-10, '2,4,6,8-tetra(C)alkyl-2,4,6,8-tetra(C)alkenylcyclotetrasiloxane,'}{'sub': 1-10', '2-10, '1,3,5-tri(C)alkyl-1,3,5-tri(C)alkenylcyclotrisiloxane,'}{'sub': 1-10', '2-10, '1,3,5,7-tetra(C)alkyl-1,3,5,7-tetra(C)alkenylcyclotetrasiloxane,'}{'sub': 1-10', '2-10, '1,3,5,7,9-penta(C)alkyl-1,3,5,7,9-penta(C)alkenylcyclopentasiloxane,'}{'sub': 1-10', '2-10, '1,3,5-tri(C)alkyl-1,3,5-tri(C)alkenylcyclotrisiloxane,'}{'sub': 1-10', '2-10, '1,3,5,7-tetra(C)alkyl-1,3,5,7-tetra(C)alkenylcyclotetrasiloxane,'}{'sub': 1-10', '2-10, '1,3,5,7,9-penta(C)alkyl-1,3,5,7,9-penta(C)alkenylcyclopentasiloxane,'}{'sub': 1-10', '2-10, '1,3,5-tri(C)alkyl-1,3,5-tri(C)alkenylcyclotrisiloxane,'}{'sub': 1-10', '2-10, '1,3,5,7-tetra (C)alkyl-1,3,5,7-tetra(C)alkenylcyclotetrasiloxane,'}{'sub': 1-10', '2-10, '1,3,5,7,9-penta(C)alkyl-1,3,5,7,9-penta(C)alkenylcyclopentasiloxane,'}{'sub': '2-10', 'hexa(C)alkenylcyclotrisiloxane,'}{'sub': '2-10', 'octa(C)alkenylcyclotetrasiloxane,'}{'sub': '2-10', 'deca(C)alkenylcyclopentasiloxane, and'}a combination thereof.5. The cell culture substrate of claim 1 , wherein the polymer formed of the ...

METHOD FOR DETECTING AND ISOLATING INVASIVE CANCER STEM CELLS EMPLOYING CELL-SURFACE AMFR AND THE USE THEREOF

The present disclosure provides an isolated invasive cancer stem cell (iCSC) or a substantially homogeneous iCSC population comprising said iCSC, which is positive for the marker AMFR on the cell membrane. The present disclosure also provides a method of detecting the iCSC within an established cancer or a collection of cancer cells. The present disclosure also provides a method of isolating the iCSC or a substantially homogeneous cell population comprising said iCSC from an established cancer or a collection of cancer cells. 126.-. (canceled)27. An isolated invasive cancer stem cell (iCSC) or a substantially homogeneous iCSC population comprising said iCSC , which is positive for the marker AMFR on the cell membrane.28. The iCSC of claim 27 , which is characterized by: (a) having stem-cell like properties; and (b) having invasive properties.29. The iCSC of claim 27 , wherein said stem-cell-like properties include: (a) expressing stem cell markers claim 27 , which comprise claim 27 , CD133 claim 27 , CD44 claim 27 , CD24 claim 27 , CD90 claim 27 , CD15 claim 27 , CD20 claim 27 , CD117 claim 27 , CD166 claim 27 , CD271 claim 27 , epithelial specific antigen (ESA) claim 27 , CXCR4 claim 27 , aldehyde dehydrogenase (ALDH) claim 27 , c-Met claim 27 , nestin claim 27 , nodal-activin claim 27 , ABCG2 claim 27 , alpha2betal-integrin claim 27 , alpha6-integrin or any combination of the foregoing; (b) expressing a low level of CD24 if said solid tumor is a breast cancer or a prostate cancer; (c) giving rise to additional stem-cell-like tumor cells; (d) being able to form a detectable tumor upon transplantation into an immunocompromised host; and/or (e) being able to regenerate the hierarchical organization of solid tumor tissues.30. The iCSC of claim 27 , wherein said cell membrane is within claim 27 , the regions of invadopodia-like structures claim 27 , said invadopodia-like structures are transient actin-based protrusions on a cancer cell that mediate focal degradation of ...

PROCESS FOR CONTINUOUS CELL CULTURE OF CANCER CELLS AND CANCER STEM CELLS

The present invention is directed towards compositions and methods of culturing cancer cells, with the methods comprising culturing cancer cells in the presence a cell culture medium while inhibiting the activity of Rho kinase (ROCK) in the cells during culturing. 126-. (canceled)27. A composition comprising fibroblast growth factor (FGF) , epithelial growth factor (EGF) , insulin growth factor-1 (IGF-1) , insulin , progesterone , transferrin , putrescine , pyruvate , albumin , selenite , thiamine , glutathione , ascorbic acid , and at least one Rho kinase (ROCK) inhibitor.28. The composition of claim 27 , wherein the composition further comprises glucose.29. The composition of claim 28 , wherein the composition further comprises at least one amino acid.30. The composition of claim 29 , wherein the composition comprises at least one amino acid selected from the group consisting of glycine claim 29 , histidine claim 29 , isoleucine claim 29 , methionine claim 29 , phenylalanine claim 29 , praline claim 29 , hydroxyproline claim 29 , serine claim 29 , threonine claim 29 , tryptophan claim 29 , tyrosine and valine.31. The composition of claim 27 , wherein the composition does not comprise animal serum.32. The composition of claim 27 , wherein the composition further comprises a base cell culture medium.33. The composition of claim 27 , wherein the ROCK inhibitor is an inhibitor of Rho kinase inhibitor 1 (ROCK 1) claim 27 , Rho kinase inhibitor 2 (ROCK 2) or both.34. The composition of claim 33 , wherein the ROCK inhibitor is selected from the group consisting of Y-27632 claim 33 , HA1100 claim 33 , HA1077 claim 33 , Thiazovivin claim 33 , and GSK429286.35. The composition of claim 33 , wherein the ROCK inhibitor is an RNA interference (RNAi) molecule specific for ROCK 1 claim 33 , ROCK 2 or both.36. A cell culture system comprising a composition according to and a culture vessel.37. The cell culture system of claim 36 , wherein the culture vessel comprises ...

THREE DIMENSIONAL MATRIX FOR CANCER STEM CELLS

Synthetic inert 3D gel culture systems are described that can be finely tuned to exhibit desired and predetermined physical, chemical, mechanical, and biochemical properties. The culture system can be utilized to study the effect of microenvironmental factors on cancer cell response, and in particular on cancer stem cell (CSC) response. Cancer cells can be encapsulated in a crosslinked gel system having a narrow range of predetermined gel stiffness. One or more biochemical factors including peptides that can affect the growth, development, and/or proliferation of CSCs can be incorporated in the system to examine the effects of the factor(s) on the encapsulated cells with regard to growth, proliferation, size, etc. 1. A method of forming a three dimensional hydrogel matrix for supporting a cancer cell , the method comprising:combining an inert synthetic polymer with a crosslinking agent to form a precursor solution;crosslinking the inert synthetic polymer via the crosslinking agent to form the three dimensional hydrogel matrix, wherein the concentration of the crosslinking agent and/or the concentration of the inert synthetic polymer is predetermined in the precursor solution such that the three dimensional hydrogel matrix has a predetermined elastic modulus; andconjugating a peptide to the matrix, the peptide affecting the growth, development, and/or proliferation of a cancer stem cell.2. The method of claim 1 , wherein the inert synthetic polymer comprises polyethylene glycol claim 1 , polyhydroxyethyl methacrylate claim 1 , polyvinylolypyrrolidone claim 1 , or polyvinyl alcohol.3. The method of claim 1 , wherein the inert synthetic polymer is combined with and reacted with the crosslinking agent prior to crosslinking the polymer.4. The method of claim 1 , further comprising encapsulating a population of cells in the three dimensional hydrogel matrix claim 1 , the population of cells comprising cancer stem cells.5. The method of claim 4 , the population of cells ...

METHODS AND REAGENTS FOR DETECTION AND TREATMENT OF ESOPHAGEAL METAPLASIA

The invention described herein relates to the treatment, detection, and diagnosis of various cancers, including esophageal or gastric adenocarcinoma and related metaplasias. The invention also includes a clonal population of Barrett's esophagus progenitor cells and methods of using them for the treatment, detection, and diagnosis of Barrett's esophagus.

CELL STRUCTURE, NON-HUMAN MODEL ANIMAL, METHOD FOR PRODUCING NON-HUMAN MODEL ANIMAL, AND METHOD FOR EVALUATING TEST SUBSTANCE

An object of the present invention is to provide a non-human model animal rich in stromal tissue, a cell structure useful for producing the above-described non-human model animal, a method for evaluating a test substance in which the above-described non-human model animal is used, and a method for producing the above-described non-human model animal. According to the present invention, a cell structure is provided which contains a biocompatible macromolecular block and at least a cancer cell and a mesenchymal cell, and in which a plurality of the above-described biocompatible macromolecular blocks are arranged in gaps between a plurality of the above-described cells. 1. A cell structure comprising:a biocompatible macromolecular block; andat least a cancer cell,wherein a plurality of the biocompatible macromolecular blocks are arranged in gaps between a plurality of the cells.2. The cell structure according to claim 1 ,wherein the cell structure comprises a mesenchymal cell.3. The cell structure according to claim 1 ,wherein the cell structure is transplanted into a non-human animal in order to produce a non-human model animal.4. The cell structure according to claim 1 ,wherein the cancer cell is an established cancer cell.5. The cell structure according to claim 1 ,wherein the cancer cell is a pancreatic cancer cell.6. The cell structure according to claim 2 ,wherein the mesenchymal cell is a mesenchymal stem cell.7. The cell structure according to claim 1 ,wherein a biocompatible macromolecule is recombinant gelatin.8. The cell structure according to claim 7 , {'br': None, 'sub': n', 'm, 'A-[(Gly-X-Y)]-B\u2003\u2003Formula, 'wherein the recombinant gelatin is represented by the following formula,'}in the formula, A represents an arbitrary amino acid or an amino acid sequence, B represents an arbitrary amino acid or an amino acid sequence, n pieces of X each independently represent any amino acid, n pieces of Y each independently represent any amino acid, n ...

ISOLATION AND USE OF SOLID TUMOR STEM CELLS

A small percentage of cells within an established solid tumor have the properties of stem cells. These solid tumor stem cells give rise both to more tumor stem cells and to the majority of cells in the tumor that have lost the capacity for extensive proliferation and the ability to give rise to new tumors. Thus, solid tumor heterogeneity reflects the presence of tumor cell progeny arising from a solid tumor stem cell. This discovery is the basis for solid tumor stem cell compositions, methods for distinguishing functionally different populations of tumor cells, methods for using these tumor cell populations for studying the effects of therapeutic agents on tumor growth, and methods for identifying and testing novel anti-cancer therapies directed to solid tumor stem cells. 1185-. (canceled)187. The isolated population of solid tumor stem cells of claim 186 , wherein said solid tumor stem cells further express B38.1.188. The isolated population of solid tumor stem cells of claim 186 , wherein said solid tumor stem cells further express epithelial specific antigen (ESA).189. The isolated population of solid tumor stem cells of claim 186 , wherein the solid tumor stem cells express low levels of CD24.190. The isolated population of solid tumor stem cells of claim 186 , wherein the solid tumor stem cells do not express CD24.192. The enriched population of claim 191 , wherein the population is at least 5-fold enriched.193. The enriched population of claim 191 , wherein the population is at least 10-fold enriched.194. The enriched population of claim 191 , wherein the population is at least 50-fold enriched.195. The enriched population of claim 191 , wherein said solid tumor stem cells further express B38.1.196. The enriched population of claim 191 , wherein said solid tumor stem cells further express epithelial specific antigen (ESA).197. The enriched population of claim 191 , wherein the solid tumor stem cells express low levels of CD24.198. The enriched population of ...

TUMOR STEM CELLS

Tumor stem cells can be obtained by culturing a tumor cell population, and exposing the cultured tumor cell population to free radicals. In certain embodiments, the free radical agent can be a nitric oxide (NO) donor. In one embodiment, the free radical agent can be Diethylenetriamine NONOate (DETA NONOate) or agents that constitutively increase cellular nitric oxide, such as phosphodiesterase inhibitors or L-arginine, or agents that increase NO synthase in the population. The methods can further include inducing stem cells present in the population to expand and/or inducing dedifferentiation of tumor cells into tumor stem cells. Additionally, the present invention provides methods of selecting stem cells from a tumor cell population. Another aspect provides methods of screening for anti-tumor stem cell teherapeutic compounds by providing high nitric oxide (HNO) tumor cells, exposing the HNO cells to at least one compound, assessing one or more indicators of HNO cell health and determining toxicity of the compound to HNO tumor cells. 1. A method of obtaining tumor stem cells comprisingculturing a cell population, andexposing the cultured cell population to free radicals to obtain the tumor stem cells.2. The method of wherein the step of exposing the cultured cell population to free radicals comprises exposing the cultured cell population to nitrogen-based free radicals.3. The method of wherein nitrogen-based free radicals includes nitric oxide (NO).4. The method of wherein nitrogen-based free radicals includes a NO donor.5. The method of wherein the step of exposing the cultured cell population further comprises exposing the population to Diethylenetriamine NONOate (DETA NONOate).6. The method of wherein the step of exposing the cultured cell population further comprises increasing NO synthase in the population.7. The method of wherein the step of exposing the cultured cell population to free radicals comprises exposing the cultured cell population to oxygen-based ...

Isolation of non-embryonic stem cells and uses thereof

The invention described herein relates to methods of isolating non-embryonic stem cell, e.g., adult stem cell, from a non-embryonic tissue, e.g., an adult tissue or organ. Non-embryonic stem cells (e.g., adult stem cells) thus isolated from the various tissues or organs can self-renew or propagate indefinitely in vitro, are multipotent and can differentiate into the various differentiated cell types normally found within the tissue or organ from which the stem cells are isolated. In addition, the isolated stem cells can be propagated through clonal expansion of a single isolated stem cell, to produce a clone of which at least about 40%, 70%, or 90% or more cells within the clone can be further passaged as single cell originated clones.

System and Method for Cold Atmospheric Plasma Treatment on Cancer Stem Cells

A method for treating cancer stem cells with cold atmospheric plasma using a cold atmospheric plasma system The method comprises the steps of placing an exit port of a cold plasma delivery device 5 cm or less from target cancer stem cells, flowing the inert gas from the source through the housing at a flow rate of 5-10 ml/minute, applying electrosurgical energy of 2-5 kV at a frequency of 20-35 kHz to at least one of the central electrode and the ring electrode to produce a cold plasma jet from the exit port, directing the cold plasma jet onto the target cancer stem cells, and applying the cold plasma jet onto the target cancer stem cells for at least 2 minutes. In a preferred embodiment the inert gas is helium. 1. A method for treating cancer stem cells with cold atmospheric plasma using a cold atmospheric plasma system having a source of high voltage power , a source of an inert gas , and a cold plasma delivery apparatus having a housing , a channel within said housing connected to said source of insert gas and having an exit port through which said inert gas exits said housing , a central electrode within said housing , and an outer ring electrode outside said housing , the method comprising the steps of:placing said exit port 5 cm or less from target cancer stem cells;flowing said inert gas from said source through said housing at a flow rate of 5-10 ml/minute;applying cold plasma jet at voltage of 2-5 kV at a frequency of 20-35 kHz to at least one of said central electrode and said ring electrode to produce a cold plasma jet from said exit port;directing said cold plasma jet onto the target cancer stem cells; andapplying said cold plasma jet onto said target cancer stem cells for at least 2 minutes.2. The method according to wherein said inert gas comprises helium.3. A method for treating cancer stem cells with cold atmospheric plasma using a cold atmospheric plasma system for producing a cold atmospheric plasma jet claim 1 , said cold atmospheric plasma system ...

Delta133P53Beta and Delta133P53Gamma Isoforms Are Biomarkers of Cancer Stem Cells

The present invention is in the field of oncology, and more particularly of cancer stem cells. It relates to a method for producing cancer stem cells based on overexpression of Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms; a method for predicting the risk that treatment with a chemotherapeutic anti-cancer agent induces cancer stem cells in a subject suffering from cancer from a cancer sample of said subject, based on detection of an increase in Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms following chemotherapeutic anti-cancer treatment; to therapeutic uses of a combination of chemotherapeutic anti-cancer agent and an agent reducing Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms expression; and also to screening methods for anti-cancer stem cells agents. 116-. (canceled)17. A method for predicting a risk of cancer metastasis in a subject suffering from cancer from a cancer sample of said subject , comprising:a) detecting sphere-forming cancer cells expressing Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms in said cancer sample; andb) concluding to the presence of a significant risk of cancer metastasis in said subject if sphere-forming cancer cells expressing Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms are detected and to the absence of a significant risk of cancer metastasis in said subject if sphere-forming cancer cells expressing Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms are not detected.18. The method according to claim 17 , wherein the subject is suffering from one or more of:(i) solid cancers selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, throat cancer, and gastrointestinal cancer including colorectal cancer oesophageal cancer, gastric cancer, pancreatic cancer, hepatocellular carcinoma, cholangiocellular cancer and teratocarcinoma, or(ii) ...

A method to up-regulate cancer stem cell markers for the generation of antigen specific cytotoxic effector t cells

The invention concerns a method of preparing a composition comprising stimulated immune system cells such as dendritic cells (DC) for use in inducing immune response of cytotoxic T lymphocytes against colorectal cancer. The dendritic cells are pulsed by contact with colorectal cancer stem cells (CSC) or their fragments thereof. These colorectal CSCs are produced by OSKM (Oct4, Sox2, Klf4 and c-Myc) induced reprogramming of differentiated colorectal cancer cells (CRC) which results in undifferentiated colorectal CSC-like cells. Both the CSC-like cells and the lysates of heat-shocked CSC-like cells could be used as an accessible source of tumour antigens for DC pulsing to induce specific immune responses against colorectal CSCs.

RNA EDITING BIOMARKERS FOR DIAGNOSIS, PHARMACOLOGICAL SCREENING AND PROGNOSTICATION IN CANCER

Compositions and methods for expanding CD34+ cells, performing research related to cancer stem cells, RNA-editing enzymes and for monitoring, diagnosing and treating, ameliorating and preventing diseases such as cancers or inflammatory diseases. 121.-. (canceled)22. A method of measuring and tracing the A-to-I RNA editing changes in a cancer stem cell(s) comprisinga) introducing into cancer stem cells a reporter vector comprising dual-luciferase, or enhanced green fluorescence protein (EGFP) or enhanced yellow fluorescence protein (EYFP) as a readout of A-to-I editing activity and a stop codon (TAG) in a hairpin structure which is part of the promoter sequence, wherein the stop codon is removed due to RNA editing resulting in a reporter gene signal as a readout of RNA editing level in which increased reporter activity correlates with higher RNA-editing activity and decreased reporter activity correlates with lower RNA-editing activity; andb) correlating changes in A-to-I RNA editing after exposure to agents.23. The method of wherein the agents are toxins claim 22 , other chemicals or biological agents.24. The method of claim 22 , wherein the cancer stem cell(s) are in an in vitro stromal co-culture system or in vivo xenograft mouse model.2533.-. (canceled)34. A method for treating claim 22 , ameliorating or preventing diseases and conditions associated with the downregulation of one or more microRNA identified in claim 22 , comprising: administering to a subject in need of treatment a composition that results in upregulation of one or more of the downregulated microRNA claim 22 , wherein the composition is one or more of antisense DNA claim 22 , RNAi claim 22 , ribozyme claim 22 , short hairpin RNA (shRNA) claim 22 , a small molecule claim 22 , an antibody or antibody fragment claim 22 , a small HA oligosaccharide claim 22 , or soluble HA-binding proteins.35. The method according to claim 34 , wherein the downregulation of the microRNA is associated with a stem cell ...

ISOLATION OF NON-EMBRYONIC STEM CELLS AND USES THEREOF

The invention described herein relates to methods of isolating non-embryonic stem cell, e.g., adult stem cell, from a non-embryonic tissue, e.g., an adult tissue or organ. Non-embryonic stem cells (e.g., adult stem cells) thus isolated from the various tissues or organs can self-renew or propagate indefinitely in vitro, are multipotent and can differentiate into the various differentiated cell types normally found within the tissue or organ from which the stem cells are isolated. In addition, the isolated stem cells can be propagated through clonal expansion of a single isolated stem cell, to produce a clone of which at least about 40%, 70%, or 90% or more cells within the clone can be further passaged as single cell originated clones. 1. A method for isolating a non-embryonic stem cell from a non-embryonic tissue , the method comprising: (a) a Notch agonist;', '(b) a ROCK (Rho Kinase) inhibitor;', '(c) a Bone Morphogenetic Protein (BMP) antagonist;', '(d) a Wnt agonist;', '(e) a mitogenic growth factor; and,', '(f) insulin or IGF (or an agonist thereof);', 'said medium optionally further comprising at least one of:', '(g) a TGFβ receptor inhibitor; and,', '(h) nicotinamide or an analog thereof;, '(1) culturing dissociated epithelial cells from the non-embryonic tissue, in contact with a first population of lethally irradiated feeder cells and a basement membrane matrix, to form epithelial cell clones, in a medium comprising(2) isolating single cells from said epithelial cell clones, and,(3) culturing isolated single cells from step (2) individually to form single cell clones, in contact with a second population of lethally irradiated feeder cells and a second basement membrane matrix in said medium;wherein each of said single cell clones represents a clonal expansion of said non-embryonic stem cell, thereby isolating said non-embryonic stem cell.24-. (canceled)5. The method of claim 1 , further comprising isolating single non-embryonic stem cell from said single ...

GENERATION OF CANCER STEM CELLS AND USE THEREOF

Methods, kits and compositions for generating cancer stem cells are provided. 1. A method for generating a cancer stem cell (CSC) , the method comprising:a. providing a differentiated cancer cell,b. incubating said differentiated cancer cell in a first medium with a pH between 5 and 6.2,c. incubating said differentiated cancer cell in a second medium supplemented with epidermal growth factor (EGF) and fibroblast growth factor (FGF),optionally further comprising analyzing expressed and secreted markers of said generated CSC, thereby generating a CSC.2. The method of claim 1 , wherein said differentiated cancer cell is selected from the group consisting of: a primary tumor cell from a subject claim 1 , a cell of a cancer cell line claim 1 , a circulating tumor cell from a subject's blood or lymph and a cancer cell differentiated in culture.3. The method of claim 1 , wherein said first medium has a pH between 5.8 and 6.2 claim 1 , optionally wherein said incubating in a first medium is performed for 15-120 minutes.4. (canceled)5. The method of claim 1 , wherein said second medium is further supplemented with a supplement comprising at least one ofa. at least one cytokine selected from the group consisting of: IL-6, TGF-β, TNF-α, CTGF, SPARC, and SDF1;b. a small molecule selected from the group consisting of: a Rho-associated protein kinase (ROCK) inhibitor, a GSK3 inhibitor, a sonic hedgehog (SHH) inhibitor, an ERK activator, a monoamine oxidase (MAO) inhibitor, a protein kinase C (PKC) activator, a histone modifying enzyme inhibitor, and a histone modifying enzyme activator; andc. a small molecule selected from the group consisting of: Y-27632, CHIR99021, tranylcypromine, PMA, 3-deazaneplanocin A, 5-aza-2′-deoxycytidine, 4-phenylbutyrate, PKC alpha activator, PKC epsilon activator, Ceramide C6, valproic acid and insulin.6. The method of claim 5 , wherein said second medium is further supplemented with TGF-β.7. The method of claim 1 , wherein the incubation with a ...

METHOD FOR CULTURING A SUBPOPULATION OF CIRCULATING EPITHELIAL TUMOUR CELLS FROM A BODY FLUID

The invention relates to a method for culturing a subpopulation of circulating epithelial tumour cells from a body fluid of a human or animal suffering from an epithelial tumour, wherein cells contained in the body fluid each containing at last one cell nucleus are separated from the body fluid and cultured over at least 24 hours in suspension, with formation of spheroids. 1. A method for culturing circulating epithelial tumor cells from a body fluid from a human or animal affected by an epithelial tumor , comprising:(i) separating cells present in the body fluid and containing at least one nucleus in each case from the body fluid without selection of certain of these cells;(ii) transferring said separated cells to a cell culture medium; and(iii) culturing said transferred separated cells under cell culture conditions in said cell culture medium containing at least an animal serum, a growth stimulator and one growth factor, wherein the growth stimulator is insulin and hydrocortisone and wherein the growth factor is Epidermal growth factor, and wherein the cells are cultured for at least 24 hours in suspension at least until a subpopulation of tumor cells, which does not require adherence to a surface for proliferation, has formed spheroids by proliferation, and wherein tumor cells which have formed the spheroids during culturing are separated from the cultured tumor cells by separating the spheroids formed during culture.2. The method as claimed in claim 1 , wherein the body fluid is lymph or blood.3. The method as claimed in claim 1 , wherein the body fluid contains intact erythrocytes and the cells containing at least one nucleus in each case are separated from the intact erythrocytes.4. The method as claimed in claim 1 , wherein the cell culture conditions comprise a temperature within the range of 35.5° C. to 37.5° C. claim 1 , and an atmosphere containing 4.5% to 5.5% CO.5. The method as claimed in claim 1 , wherein fresh cell culture medium is added to the ...

Fluorescent Synthetic Retinoids

There are described novel compounds of formula I: which R, R, R, R, R, R, R, R, Rand Rare each as herein defined. 2. A compound according to in which Ris a group II , as defined in .3. A compound according to in which Ris a group III , as defined in .4. A compound according to in which Ris a group IV , as defined in .5. A compound according to in which Ris a group V , as defined in .6. A compound according to in which Ris a group VI , as defined in .7. A compound according to in which Ris a group VII , as defined in .8. A compound according to in which Ris a group VIII , as defined in .9. A compound according to in which Ris a group IX , as defined in .10. A compound according to in which Ris a group X , as defined in .11. A compound according to in which Ris a group XI , as defined in .12. A compound according to in which the moiety —CORis in the 4-position.13. A compound according to in which the moiety —CORis in the 3-position.14. A compound according to in which Ris hydrogen.15. A compound according to wherein at least 60% by weight of the compound remains following 3 days exposure to light having a wavelength of 300 to 400 nm.16. A compound according to for use in the control of cell differentiation or apoptosis.17. A compound according to claim 1 , which is selected from the group consisting of:4-2-[4,4-dimethyl-1-(propan-2-yl)-1,2,3,4-tetrahydroquinolin-6-yl]ethynylbenzoic acid (9); and6-(1,4,4-trimethyl-1,2,3,4-tetrahydroquinolin-6-yl)-naphthalene-2-carboxylic acid methyl ester (11);3-[4-(1,4,4-trimethyl-1,2,3,4-tetrahydroquinolin-6-yl)-phenyl]-acrylic acid methyl ester (13); and4-2-[2,4,4-trimethyl-1-(propan-2-yl)-1,4-dihydroquinolin-6-yl]ethynylbenzoic acid, (17);and isomers thereof;in free or in salt form.17. (canceled)1821.-. (canceled)22. A method of inducing the differentiation of a stem cell comprising the steps of:(i) forming a preparation of stem cells in a cell culture medium suitable for maintaining said stem cells wherein said culture medium ...

Isolated liver cancer stem cell and method for drug screening thereof

An isolated rat liver cancer stem cell line which is named as TW-1 is provided. A method for drug screening by using the isolated rat liver cancer stem cell line is also provided.

CULTURE MEDIUM AND METHOD FOR ENRICHING AND MAINTAINING CANCER STEM CELLS (CSCS) USING SAID MEDIUM

The present invention relates to a serum-free conditioned medium that solves the drawbacks mentioned in the prior art, as it does not require prior handling of the cells, and it furthermore allows starting from a large population with no additional cost. This medium favors in vitro proliferation and conservation of the pluripotency potential that allows maintaining a state that is undifferentiated with respect to the subpopulation of cancer stem cells (CSCs) and in turn does not allow survival of the differentiated cells. 1. A cell culture medium suitable for isolating and/or enriching cancer stem cells (CSCs) , obtained or obtainable by a method comprising the following steps:a. Seeding mesenchymal stem cells (MSCs) in a plate, vessel or flask suitable for culturing cells using a culture medium suitable for this purpose;b. Optionally removing any remainder of medium and/or serum from the culture medium from step a) by means of washing;c. Adding a sphere medium or conventional medium without FBS to the product of step b);d. Collecting the culture medium from step c) and adding new sphere medium;e. Repeating the process from step d) until the cells in culture reach a level of confluence of 80-90%;f. Optionally filtering and freezing the medium obtained in steps d) and e) until use.2. The cell culture medium according to claim 1 , wherein said medium is obtained or obtainable by a method comprising the following steps:a. Seeding mesenchymal stem cells in a plate, vessel or flask suitable for culturing cells using a conventional culture medium, where said cells have a confluence between 40% and 90% at the start of the process;b. Removing any remainder of medium and/or serum from the culture medium from step a) by means of washing at least 24 hours after the start of the process;c. Adding a sphere medium or conventional medium without FBS to the product of step b);d. Collecting the culture medium from step c) at least 48 hours after adding the medium in step c), and ...

Culture Medium for Expanding Breast Epithelial Stem Cells

The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids.

Rapid Method Production High Purity Cancer Stem Cells and Population of High Purity Cancer Stem Cells

The disclosure provides reagents, including cells, and related methods, useful for administering to subjects with a neoplastic disorder. The reagents and methods encompass cancer stem cells of enhanced purity. Neoplastic disorder encompasses melanoma, ovarian cancer, colorectal cancer, breast cancer, and lung cancer. 125-. (canceled)26. A method for producing purified adherent melanoma cancer stem cells , comprising the steps of:(a) obtaining a tissue specimen from a biopsy of a melanoma tumor from a subject;(b) dissociating the tissue specimen mechanically;(c) enzymatically dissociating samples of the tissue specimen into a single-cell suspension(d) immersing the single-cell suspension of (c) in serum free cell culture media comprising basic fibroblast growth factor (bFGF) and culturing on a low adherent or an ultra-low adherent surface to produce a cancer cell culture comprising melanoma cancer cell spheroids;(e) sedimenting the spheroids to collect microspheres;(f) dissociating cells from the microspheres to yield a single-cell suspension;(g) transferring the single-cell suspension of (f) to an adherent substrate and expanding cell number in vitro to establish a population of purified adherent melanoma cancer stem cells comprising CD146+/CD271−, CD146+/CD271+, and CD146−/CD271+ cells.2730.-. (canceled)31. The method according to claim 26 , wherein the population of purified adherent cancer stem cells comprises at least 20% claim 26 , at least 30% claim 26 , at least 40% claim 26 , at least 50% claim 26 , at least 60% claim 26 , at least 70% claim 26 , at least 80% claim 26 , at least 90% claim 26 , at least 95% claim 26 , or at least 98% cells expressing CD146.32. The method according to claim 26 , wherein the population of purified adherent cancer stem cells comprises at least 20% claim 26 , at least 30% claim 26 , at least 40% claim 26 , at least 50% claim 26 , at least 60% claim 26 , at least 70% claim 26 , at least 80% claim 26 , at least 90% claim 26 , at ...

CANCER-ASSOCIATED FIBROBLASTS IN MAINTAINING STEMNESS OF CANCER STEM CELLS

An in vitro co-culture system comprising cancer-associated fibroblasts (CAFs) and cancer cells for producing and maintaining cancer stem cells and uses thereof for identifying agents capable of reducing cancer cell stemness. Also disclosed herein are a paracrine network through which CAFs facilitate production and/or maintenance of cancer stem cells and the use of components of such a paracrine network for prognosis purposes and for identifying cancer patients who are likely to respond to certain treatment. 1. An in vitro co-culture system , comprising cancer associated fibroblasts (CAFs) and a population of cancer cells , wherein the CAFs are CD90 , and wherein the CAFs maintain stemness of the cancer cells in the co-culture system.2. The in vitro co-culture system of claim 2 , wherein the CAFs are obtained from a cancer patient.3. The in vitro co-culture system of claim 3 , wherein the cancer patient is a lung cancer patient claim 3 , a breast cancer patient claim 3 , a kidney cancer patient claim 3 , a prostate cancer patient claim 3 , an ovary cancer patient claim 3 , a skin cancer patient claim 3 , a cervical cancer patient claim 3 , a colon cancer patient claim 3 , a liver cancer patient claim 3 , a melanoma patient claim 3 , an oral cancer patient claim 3 , or a pancreatic cancer patient.4. The in vitro co-culture system of claim 3 , wherein the cancer patient is a non-small cell lung cancer patient.5. The in vitro co-culture system of claim 1 , wherein the population of cancer cells include lune cancer cells claim 1 , breast cancer cells claim 1 , kidney cancer cells claim 1 , prostate cancer cells claim 1 , ovary cancer cells claim 1 , skin cancer cells claim 1 , cervical cancer cells claim 1 , colon cancer cells claim 1 , liver cancer cells claim 1 , melanoma cells claim 1 , oral cancer cells claim 1 , or pancreatic cancer cells.6. The in vitro co-culture system of claim 5 , wherein the cancer cells are non-small cell lung cancer cells.7. The in vitro co- ...

METHODS AND PRODUCTS FOR TRANSFECTION

The present invention relates in pan to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed. 19.-. (canceled)10. A method for reprogramming a differentiated cell to a less differentiated state , comprising:(a) providing a differentiated cell;(b) culturing the differentiated cell;(c) transfecting the differentiated cell with one or more synthetic RNA molecules, wherein the one or more synthetic RNA molecules include at least one RNA molecule encoding one or more reprogramming factors and wherein the transfecting results in the cell expressing the one or more reprogramming factors; and(d) repeating step (c) at least twice during 5 consecutive days, wherein the amount of one or more synthetic RNA molecules transfected in one or more later transfections is greater than the amount transfected in one or more earlier transfections, to result in the cell being reprogrammed to a less differentiated state,wherein steps (c) and (d) occur in the presence of a medium containing ingredients that support reprogramming of the differentiated cell to a less differentiated state.11. The method of claim 10 , wherein the differentiated cell is derived from a biopsy.12. The method of claim 10 , wherein the differentiated cell is from a human subject.13. The method of claim 11 , wherein the differentiated cell is derived from a dermal punch biopsy sample.14. The method of claim 10 , wherein the differentiated cell is a skin cell.15. The method of claim 10 , further comprising contacting the cell with at least one member of the group: poly-L-lysine claim 10 , poly-L-ornithine claim 10 , RGD peptide claim 10 , fibronectin claim 10 , vitronectin claim 10 , collagen claim 10 , and laminin.16. The method of claim 10 , wherein the synthetic RNA molecule contains at least one of a ...

METHODS AND PRODUCTS FOR TRANSFECTION

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed. 19.-. (canceled)10. A method for reprogramming a differentiated cell to a less differentiated state , comprising:(a) providing a differentiated cell;(b) culturing the differentiated cell; and wherein the one or more synthetic RNA molecules include at least one RNA molecule encoding one or more reprogramming factors;', 'wherein the transfecting results in the cell expressing the one or more reprogramming factors; and', 'wherein step (c) is performed at least twice and the amount of one or more synthetic RNA molecules transfected in one or more later transfections is greater than the amount transfected in one or more earlier transfections to result in the cell being reprogrammed to a less differentiated state and occurs in the presence of a medium containing ingredients that support reprogramming of the differentiated cell to a less differentiated state., '(c) transfecting the differentiated cell with one or more synthetic RNA molecules,'}11. The method of claim 10 , wherein the differentiated cell is derived from a biopsy.12. The method of claim 11 , wherein the differentiated cell is derived from a dermal punch biopsy sample.13. The method of claim 10 , wherein the differentiated cell is from a human subject.14. The method of claim 10 , wherein the differentiated cell is a skin cell.15. The method of claim 10 , further comprising contacting the cell with at least one member of the group: poly-L-lysine claim 10 , poly-L-ornithine claim 10 , RGD peptide claim 10 , fibronectin claim 10 , vitronectin claim 10 , collagen claim 10 , and laminin.16. The method of claim 10 , wherein the one or more synthetic RNA molecules contain at least one of a pseudouridine or a 5- ...

BIOREACTOR SYSTEM

A three dimensional cell culture and bioreactor system is provided. The system comprises one or more cell culture chamber. Each cell culture chamber comprises an inlet port and an outlet port in fluid communication with the cell culture chamber. The cell culture chambers may be segregated or in fluid communication with one another. The systems may be used to conduct drug efficacy test, isolate certain cell types from a complex tissue sample of multiple cell types, allow for the ex vivo culturing of patient tissue samples to help guide the course of treatment, and conduct co-culture experiments. 129-. (canceled)30. A method for isolating residual cell populations from tissue samples after exposure to pharmaceutical agents comprising:culturing a primary cell population derived from at least a portion of a biological sample in a cell culture chamber of a 3D cell culturing system, wherein the primary cell population comprises one or more cell types;treating the primary cell population with a first dose of one or more pharmaceutical agents, wherein exposure to the one or more pharmaceutical agents results in an increased ratio of a first residual cell type relative to other cell types in the primary cell population;culturing the residual cell type;wherein the cell culture chamber of the 3D cell culturing system comprises an inlet port and an outlet port that are in communication with an interior volume of the cell culture chamber, the interior volume of the cell culture chamber housing a cell-scaffold construct and the target cell population.31. The method of claim 30 , further comprising treating the first residual cell type with a second dose of the one or more pharmaceutical agents to obtain an enriched residual cell type.32. The method of claim 31 , further comprising:treating the cultured residual cell type with a second dose of the one or more pharmaceutical agents, or a second dose with one or more different pharmaceutical agents; anddetermining an efficacy of the ...

A NOVEL INVADOPODIA-SPECIFIC MARKER OF INVASIVE CANCER STEM CELLS AND THE USE THEREOF

The present invention provides an invasive cancer stem cell (iCSC) or a substantively homogeneous cell population including said iCSC based on a cell-surface biomarker specifically localizing to the invadopodia of said iCSC. The present invention further provides an invasive leukemia stem cell (iLSC) or a substantively homogeneous cell population including said iLSC based on a cell-surface biomarker specifically localizing to the invadopodia of said iLSC. The present invention also provides a method or a kit of determining a diagnosis of aggressive solid tumor or hematopoietic cancer. 1. An invasive cancer stem cell comprising marker PPH , wherein the marker PPH is on surface of the invasive cancer stem cell.2. The invasive cancer stem cell of claim 1 , which is characterized by: (a) having stem-cell like properties; and (b) having invasive properties.3. The invasive cancer stem cell of claim 2 , wherein the stem-cell like properties include: (a) expressing stem cell markers claim 2 , which comprise claim 2 , but not limited to claim 2 , CD133 claim 2 , CD44 claim 2 , CD24 claim 2 , CD90 claim 2 , CD15 claim 2 , CD20 claim 2 , CD117 claim 2 , CD166 CD271 claim 2 , epithelial specific antigen claim 2 , CXCR4 claim 2 , aldehyde dehydrogenase claim 2 , c-Met claim 2 , nestin claim 2 , nodal-activin claim 2 , ABCG2 claim 2 , alpha2beta1-integrin alpha6-integrin or any combination of the foregoing; (b) expressing a low level of CD24 if said solid tumor is a breast cancer or a prostate cancer; (c) giving rise to additional stem-cell-Like tumor cells; (d) being able to form a detectable tumor upon transplantation into an immunocompromised host; and/or (e) being able to regenerate the hierarchical organization of solid tumor tissues.4. A method of detecting an invasive cancer stem cell comprising marker PPH claim 2 , wherein the marker PPH is on surface of the invasive cancer stem cell claim 2 , within a solid tumor claim 2 , wherein the method comprises: (a) preparing a ...

SYSTEM FOR IMMUNOTHERAPY TARGETING TUMOR PROPAGATION AND PROGRESSION

Finding biologically relevant cancer markers is key to developing an effective treatment. Once specific antigens have been identified that are present on the cell population responsible for propagating tumors, T cells can be genetically altered to target these antigens and then used for personalized T cell therapy. A method to identify and select peptide antigens that effectively associate with and are presented by host HLA surface molecules originating from tumor cells responsible for the persistence and propagation of a cancer has been developed. The system of using these data to produce T cells engineered to express T cell receptors recognizing the peptide antigens is used in the production of a personalized adoptive T cell therapy for cancer that eliminates the cells capable of tumor propagation and cancer progression. The system is especially useful in the production of cancer treatments to achieve complete durable remission of cancers of epithelial origin. 1. A method of identifying C-RC cell-specific T cells , comprising(a) obtaining a tumor sample from an individual;(b) cultivating the tumor sample under conditions that induce a stress response in differentiating and differentiated cells but permits C-RC cells to propagate and which activate a regenerative response;(c) isolating the dominant actively expanding, most rapidly dividing population of cells from step (b); and(d) culturing the cells to 60 to 95% confluence to obtain a population of 51% to 100% C-RC{'sup': '+', 'mixing C-RC cells from step (d) with isolated CD8T cells from normal or cancer donors,'}isolating T cells that react with the treated C-RC cells, wherein the reactive T cells comprise T cell receptors that target a C-RC-specific antigen.2. The method of claim 1 , comprising treating the C-RC cells with an interferon gamma agonist to enhance expression of HLA-peptide complexes.3. The method of further comprising adding between initial lymphocyte isolation and the selection method an in vitro ...

THERAPEUTIC COMPOSITION FOR TREATMENT OF GLIOBLASTOMA

The present invention is directed to compositions and methods for treating an animal diagnosed with Glioblastoma multiforme (GBM). 1. A method of eliciting an immune response in an animal in need thereof , comprising administering to the animal an anti-tumor composition comprising an adherent glioblastoma GBM6-AD stem cell , wherein the adherent glioblastoma GBM6-AD stem cell is ATCC® Patent Deposit Designation PTA-11498.2. The method of claim 1 , further comprising administering Imiquimod to the animal.3. The method of claim 2 , wherein the Imiquimod is administered topically claim 2 , intradermally claim 2 , subcutaneously claim 2 , and/or via intralymph node injections.4. The method of claim 1 , wherein the anti-tumor composition is administered parenterally claim 1 , orally or intranasally.5. The method of claim 1 , wherein the anti-tumor composition is administered at more than one time point.6. The method of claim 1 , wherein the anti-tumor composition is administered at a dose of 100 claim 1 ,000-100 million GBM6-AD cells at each administration time point.7. The method of claim 1 , further comprising administering irradiation therapy to the animal.8. The method of claim 7 , wherein the irradiation therapy is administered at a dose of less than 6 claim 7 ,000 cGY.9. The method of claim 7 , further comprising administering a radiation sensitizer to the animal.10. The method of claim 9 , wherein the radiation sensitizer is a parp inhibitor or temozolomide.11. The method of claim 1 , wherein the animal is a mammal.12. The method of claim 1 , further comprising administering dexamethasone therapy to the animal.13. The method of claim 12 , wherein the dexamethasone therapy is administered at the time of diagnosis.14. The method of claim 1 , further comprising administering a chemotherapy or drug that depletes regulatory T cell or myeloid derived suppressor cells.15. The method of claim 14 , wherein the chemotherapy is sunititib claim 14 , ontak claim 14 , ...

COMPOSITIONS AND METHODS FOR TREATING CANCER AND BIOMARKERS TO DETECT CANCER STEM CELL REPROGRAMMING AND PROGRESSION

In alternative embodiment, provided are methods and compositions for treating, ameliorating or preventing diseases and conditions, such as cancer, including cancers associated with stem cells such as, without limitation, myelodysplastic syndrome (MDS) and a myeloproliferative neoplasm like chronic myeloid leukemia (CML) or acute myeloid leukemia (AML), and ablating or killing cancer stem cells. In alternative embodiment, provided are a new set of biomarkers to detect leukemia stem cell reprogramming and CML progression. In alternative embodiment, provided are therapeutic targets for treating myelodysplastic syndrome (MDS) and chronic myeloid leukemia (CML) by targeting edited let-7 transcripts. 1. A method for:treating, ameliorating, stopping or slowing the progression of, or preventing a cancer or a cancer associated with a stem cell,inhibiting, decreasing or slowing the progression of a therapeutically responsive or a drug responsive cancer to a therapeutically resistant (drug resistant) cancer,inhibiting, decreasing or slowing the generation of self-renewing leukemia stem cells (LSCs) or the maintenance of LSCs,decreasing or inhibiting myelodysplastic syndrome (MDS) or a myeloproliferative neoplasm (MPN) initiation and/or maintenance in inflammatory microenvironments,inhibiting or decreasing the amount of GSK3β missplicing and increasing degradation of β-catenin, and/orenhancing let-7 microRNA (miRNA) biogenesis, decreasing adenosine-to-inosine (A-to-I) editing of polycistronic let-7 loci, and/or increasing levels of mature let-7 microRNA (miRNA) levels, comprising: (i) breakpoint cluster region protein (BCR)-Abelson murine leukemia viral oncogene homolog 1 (ABL1), or BCR-ABL1 (a BCR-ABL fusion protein),', '(ii) double-stranded RNA-specific adenosine deaminase ADAR1), or', '(iii) ADAR1 and BCR-ABL1; or, '(a) administering to a subject in need thereof, or in need of treatment, an agent or combination of agents that inhibit or decrease the expression or activity of ...

Transformed human pluripotent stem cells and associated methods

The present disclosure provides transformed human pluripotent stem cell (t-hPSC). t-hPSCs are not dependent on Oct4 for renewal and survival, however exhibit a sensitivity to reduced levels of the transcription factor Nanog. Also provided are methods of culturing cells for use in a cell-based screening assay comprising placing one or more transformed human pluripotent stem cells into a receptacle and culturing said stem cells in the receptacle to form a monolayer of stem cells without cell overlap. Methods of screening compounds using t-hPSCs are also described.

Methods for Generation of Cytocapsulae and Cytocapsular Tubes

This invention provides for methods and compositions for generation of cytocapsulae and cytocapsular tubes in a 3D matrix. 1. A composition comprising a 3D matrix and cytocapsulae and cytocapsular tubes.2. The composition of wherein the cytocapsulae and cytocapsular tubes are generated by a single cell implanted at the top of the 3D matrix.3. The composition of wherein the cytocapsulae and cytocapsular tubes comprise the cell that generates the cytocapsulae and cytocapsular tubes claim 1 , or comprise multiple cells that enter into the cytocapsulae and cytocapsular tubes from surrounding environment claim 1 , or are devoid of cell after ecellularization.45.-. (canceled)6. The composition of wherein the 3D matrix is biodegradable.7. The composition of wherein the 3D matrix comprises elastin claim 1 , laminin claim 1 , collagen claim 1 , proteoglycans claim 1 , and/or non-proteoglycan polysaccharide.8. The composition of wherein the proteoglycan comprises heparan sulfate claim 7 , chondroitin sulfate claim 7 , and keratin sulfate.9. The composition of wherein the non-proteoglycan polysaccharide comprises hyaluronic acid.10. The composition of wherein the 3D matrix further comprises fibrillary protein claim 1 , Facit protein claim 1 , short chain protein and basement membrane protein.11. The composition of wherein the 3D matrix further comprises signal proteins comprising focal adhesion kinase (FAK) claim 1 , talin claim 1 , vinculin claim 1 , paxllin claim 1 , α-actinin claim 1 , and GTPase.12. The composition of wherein the 3D matrix is a Matrigel.13. The composition of wherein polymerization claim 1 , density claim 1 , protein concentration claim 1 , and viscoelasiticity of the 3D matrix affect the generation of the cytocapsulae and cytocapsular tubes.14. A method of generating cytocapsulae and cytocapsular tubes comprising:implanting cells on top of a 3D matrix, andincubating the cell implanted 3D matrix under conditions such that the cells engender the ...

METHODS AND MATERIALS FOR HEMATOENDOTHELIAL DIFFERENTIATION OF HUMAN PLURIPOTENT STEM CELLS UNDER DEFINED CONDITIONS

Methods and compositions for differentiating pluripotent stem cells into cells of endothelial and hematopoietic lineages are disclosed. 115-. (canceled)16. A method of treating a blood disorder in a subject , the method comprising administering therapeutic cells comprising CD43 hematopoietic progenitors composed of CD43CD235aCD41a erythromegakaryocytic progenitors and lin−CD34CD43CD45 multipotent hematopoietic progenitors to treat the blood disorder , the therapeutic cells made by the method comprising the steps of:{'sup': EMH', '−', '+', '+', '+, '(a) exposing human pluripotent stem cells to a xenogen-free and serum albumin-free mixture comprising components of about 25 ng/ml to about 50 ng/ml FGF2, at least 50 ng/ml to about 250 mg/ml of bone morphogenetic protein 4 (BMP4), Activin A of less than 15 ng/ml, and about 1 mM to about 2 mM LiCl under hypoxic conditions for a period of about two days to form a population of therapeutic linKDRAPLNRPDGFRalpha primitive mesoderm cells with mesenchymoangioblast potential without the formation of embryoid bodies or coculture with stromal cell lines;'}{'sup': EMH', '+', '−', '+', 'EMH', '−', 'hi', '+', 'lo/−, '(b) exposing cells at the primitive mesoderm stage of step (a) to a mixture comprising components FGF2 and VEGF under hypoxic conditions for a period of about 1-2 days to obtain a population comprising lin−KDRAPLNRPDGFRalpha primitive mesoderm with hemangioblast (HB-CFC) potential and hematovascular mesoderm cells (linKDRAPLNRPDGFRalpha) enriched in cells with a potential to form hematoendothelial clusters when cultured on OP9 cells;'}{'sup': +', '+', '−', '+', '−', '−', '+', '−', '+', '−', '+', '+, '(c) exposing the cells at the hematovascular mesoderm stage of step (b) to a mixture comprising components FGF2, VEGF, IL6, SCF, TPO, and IL3 for about one day to achieve formation of CD144CD73CD235a/CD43 non-hemogenic endothelial progenitors (non-HEP), CD144CD73CD235a/CD43 hemogenic endothelial progenitors (HEPs), ...

METHODS AND PRODUCTS FOR TRANSFECTION

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.

BINDING PROTEINS SPECIFIC FOR LOX1 AND USES THEREOF

This disclosure provides LOX1 (LOX1) binding proteins such as anti-LOX1 antibodies, and compositions and methods for making these binding proteins. In certain aspects the LOX1-binding proteins provided herein, inhibit, or antagonize LOX1 activity. In addition, the disclosure provides compositions and methods for diagnosing and treating conditions associated with atherosclerosis, thrombosis, coronary artery disease (CAD), ischemia (e.g., myocardial ischemia), infarction (e.g., myocardial infarction), acute coronary syndrome (ACS), stroke, reperfusion injury, restenosis, peripheral vascular disease, hypertension, heart failure, inflammation (e.g., chronic inflammation), angiogenesis, preeclampsia, cancer and other LOX1-mediated diseases and conditions. 1. An isolated LOX1-binding protein comprising a heavy chain variable region (VH) having at least 90 , 95 , 97 , 98 or 99% sequence identity to SEQ ID NOs:4 , 19-29 , 41 , or 48-54; and a light chain variable region (VL) having at least 90 , 95 , 97 , 98 or 99% sequence identity to SEQ ID NOs:33 , 36 , 37 , 58 or 65-70.2. An isolated LOX1-binding protein comprising a heavy chain variable region (VH) selected from the group consisting of: a VH comprising SEQ ID NO:4 , 19-29 , 41 , or 48-54 and a light chain variable region (VL) selected from the group consisting of a VL comprising SEQ ID NO:33 , 36 , 37 , 58 or 65-70.4. An isolated LOX1-binding protein that binds the same epitope as an isolated LOX1-binding protein of .5. An isolated LOX1-binding protein which competes for binding to LOX1 with an isolated LOX1-binding protein of .6. The isolated LOX1-binding protein of claim 3 , wherein the LOX1-binding protein is an antibody.7. The isolated LOX1-binding protein of claim 6 , wherein the antibody is a monoclonal antibody claim 6 , a recombinant antibody claim 6 , a human antibody claim 6 , a humanized antibody claim 6 , a chimeric antibody claim 6 , a bi-specific antibody claim 6 , a multi-specific antibody claim 6 , or a ...

Methods and products for transfection

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.

CULTURE MEDIUM

The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids. 1. A method for expanding epithelial stem cells comprising:providing a population of epithelial stem cells;providing a culture medium comprising an ErbB3/4 ligand, a receptor tyrosine kinase ligand and a BMP inhibitor;contacting the stem cells with the culture medium; andculturing the cells under appropriate conditions.2. The method of claim 1 , wherein the culture medium further comprises a Wnt agonist.3. The method of or claim 1 , wherein the ErbB3/4 ligand is a neuregulin polypeptide.4. A culture medium comprising a receptor tyrosine kinase ligand and a BMP inhibitor claim 1 , characterised in that the culture medium further comprises an ErbB3/4 ligand.5. The culture medium of claim 4 , wherein the culture medium further comprises a Wnt agonist.6. The culture medium of or claim 4 , wherein the ErbB3/4 ligand is a neuregulin polypeptide.7. The culture medium of claim 6 , wherein the neuregulin polypeptide comprises or consists of the amino acid sequence recited in SEQ ID NO: 27.8. The culture medium of any one of - claim 6 , wherein the receptor tyrosine kinase ligand is selected from EGF claim 6 , HGF claim 6 , PDGF and FGF (e.g. FGF7 and/or FGF10).9. The culture medium of any one of - claim 6 , wherein the Wnt agonist is an Lgr5 agonist.10. The culture medium of claim 9 , wherein the Lgr5 agonist is Rspondin.11. The culture medium of any one of - claim 9 , wherein the BMP inhibitor is Noggin.12. The culture medium of any one of - claim 9 , wherein the culture medium further comprises a TGF-beta inhibitor.13. The culture medium of any one of - claim 9 , wherein the culture medium further comprises: (i) a Notch inhibitor and/or a prostaglandin pathway activator claim 9 , and/or (ii) a cAMP pathway activator and/or a BMP pathway activator claim 9 , and/or (iii) a p38 inhibitor claim ...

METHOD FOR MANUFACTURING THREE-DIMENSIONAL CELL CULTURE SUPPORT HAVING DOUBLE CROSSLINK, AND CASTING TRAY FOR MANUFACTURING THREE-DIMENSIONAL CELL CULTURE SUPPORT

The present disclosure relates to a method for manufacturing a three-dimensional cell culture support having a double crosslink, and a casting tray for manufacturing the three-dimensional cell culture support, wherein the method for manufacturing the three-dimensional cell culture support having the double crosslink includes: producing a cell mixed hydrogel; manufacturing a casting gel mold in a three-dimensional shape; and manufacturing a structure gelated in a three-dimensional shape, and the casting tray for manufacturing the three-dimensional cell culture support includes: a tray part including a groove accommodating a gel solution; a mold part covering the tray part; and a mold protrusion provided on the mold part and inserted into the groove when the mold part covers the tray part. 1. A method for manufacturing a three-dimensional cell culture support having a double crosslink , comprising:producing a cell mixed hydrogel;manufacturing a casting gel mold in a three-dimensional shape; anddispersing the cell mixed hydrogel into the casting gel mold manufactured in the three-dimensional shape and gelating the cell mixed hydrogel to manufacture a structure gelated in a three-dimensional shape.2. The method of claim 1 , wherein the producing of the cell mixed hydrogel comprises:mixing gelatin with alginate to prepare a mixed solution;filtering the mixed solution to produce a hydrogel; andmixing the hydrogel with cells.3. The method of claim 2 , wherein the mixed solution is one selected from a group consisting of apatite claim 2 , cellulose claim 2 , gellan claim 2 , agarose claim 2 , chitosan claim 2 , keratin claim 2 , and collagen claim 2 , or claim 2 , a combination of two or more of apatite claim 2 , cellulose claim 2 , gellan claim 2 , agarose claim 2 , chitosan claim 2 , keratin claim 2 , and collagen.4. The method of claim 2 , wherein the mixed solution is one selected front a group consisting of a transforming growth factor (TGF) claim 2 , a vascular ...

METHOD AND CULTURE MEDIUM FOR EX VIVO CULTURING OF EPIDERMIS-DERIVED STEM CELLS

The present invention relates to a method for culturing epidermis-derived stem cells comprising the step of culturing epidermis-derived stem cells in the presence of a three-dimensional extracellular matrix (3D-ECM) and a basal cell culture medium comprising: Epidermal Growth Factor (EGF); and/or a Vascular Endothelial Growth Factor (VEGF); and/or a Fibroblast Growth Factor (FGF); and further a ROCK (Rho-kinase) inhibitor. The present invention further relates to a method for ex vivo de novo generation of epidermis-derived stem cells. Furthermore, the present invention relates to an epidermis-derived stem cell that is obtainable by a method according to the present invention. Uses of said epidermis-derived stem cell, e.g. uses of said epidermis-derived stem cell for in vitro tissue production, in vitro drug discovery screening and medical applications, are also provided herein. The present invention further relates to a cell culture medium that is employed in the context of a method of the present invention. 2. The method of claim 1 , wherein said basal cell culture medium comprises said EGF claim 1 , said VEGF claim 1 , said FGF and said ROCK inhibitor.3. The method of or claim 1 , wherein said Vascular Endothelial Growth Factor (VEGF) is selected from the group consisting of VEGF-164 claim 1 , VEGF-165 claim 1 , VEGF-120 and VEGF-121.4. The method of any one of to claim 1 , wherein said Fibroblast Growth Factor (FGF) is selected from the group consisting of FGF-2 claim 1 , FGF-7 claim 1 , FGF-10 and FGF-18.5. The method of any one of to claim 1 , wherein said ROCK inhibitor is selected from the group consisting of (S)-(+)-2-methyl-1-[(4-methyl-5-isoquinolinyl)sulfonyl]-hexahydro-1H-1 claim 1 ,4 diazepine dihydrochloride (H-1152) and (R)-(+)-trans-4-(1-aminoethyl)-N-(4-Pyridyl)cyclohexanecarboxamide dihydrochloride monohydrate (Y-27632).6. The method of any one of to claim 1 , wherein said basal cell culture medium further comprises a Sonic Hedgehog (SHH) inhibitor ...

METHOD FOR DELIVERING EXOGENOUS MITOCHONDRIA INTO CELLS

The present invention relates to a method for delivering exogenous mitochondria into cells and, more specifically, to a method for efficiently delivering, into the cytoplasm of target cells to be injected, mitochondria isolated from donor cells. 1. A method for delivering exogenous mitochondria into recipient cells , comprising:a) a step of mixing the recipient cells with mitochondria isolated from donor cells; andb) a step of centrifuging the resulting mixture.2. The method of claim 1 , wherein claim 1 , in the step a) claim 1 , the mitochondria and the recipient cells are present in the mixture such that the mitochondria are contained in a weight of 0.005 to 25 μg per 1×10cells of the recipient cells.3. The method of claim 1 , wherein in the step b) claim 1 , the centrifugation is performed at a temperature of 0° C. to 40° C. for 0.5 to 20 minutes with 1 to 2 claim 1 ,400×g.4. The method of claim 1 , further comprising: a step of adding a surfactant before the step b).5. The method of claim 4 , wherein the surfactant in the mixture has a concentration of 1 and 100 mg/ml.6. The method of claim 1 , further comprising: a step of performing incubation before the step b).7. The method of claim 6 , wherein the incubation is performed at a temperature of 0° C. to 40° C. for 0.1 to 4 hours.8. The method of claim 1 , wherein the donor cells and the recipient cells are respectively any one selected from the group consisting of somatic cells claim 1 , stem cells claim 1 , cancer cells claim 1 , and combinations thereof.9. The method of claim 8 , wherein the somatic cells are any one selected from the group consisting of muscle cells claim 8 , hepatocytes claim 8 , fibroblasts claim 8 , epithelial cells claim 8 , neurons claim 8 , adipocytes claim 8 , osteocytes claim 8 , leukocytes claim 8 , lymphocytes claim 8 , mucosal cells claim 8 , and combinations thereof.10. The method of claim 1 , wherein the donor cells are allogeneic or xenogeneic to the recipient cells.11. The ...

Method for the Isolation for Mammalian Stem Cells and Uses Thereof

The present invention concerns the field of stem cell biology, and in particular relates to a method for producing an isolated bona fide population of mammalian stem cells, and uses of the stem cells thus produced. Human glioblastomas (hGBMs) have now been shown to contain a minor subset of cells bearing the defining features of somatic stem cells (SCs) and the ability to establish, expand and perpetuate these tumors. They are defined stem-like tumor propagating cells (TPCs). This has caused a paradigmatic shift in the way we interpret hGBM physiology, for it identifies TPCs as a major culprit to be tackled for the development of novel therapeutics. It also suggests that studying the regulatory mechanisms of normal neurogenesis may point to specific inhibitors of TPCs. 1. A method for producing an isolated population of bona fide mammalian stem cells comprising the steps of:a. providing a population of cells,b. selecting from the population of cells of step a. the cells that express EphA2;c. isolating the cells selected in step b.,thereby producing a population of mammalian stem cells.2. The method according to claim 1 , wherein said population of cells is obtained from a tissue sample claim 1 , preferably from a biopsy claim 1 , preferably from a brain biopsy.3. The method according to or claim 1 , wherein said selection step b. is carried out by cell sorting claim 1 , preferably by Fluorescence-activated cell sorting.4. The method according to anyone of to claim 1 , wherein said selection step b. allows to sort the cells according to their EphA2 expression level.5. The method according to claim 4 , wherein said EphA2 expression level is measured with an anti-EphA2 antibody.6. The method according to anyone of to claim 4 , wherein the mammalian stem cells isolated in step c. are selected from the group consisting of human and mouse bona fide stem cells.7. The method according to anyone of claim 6 , wherein the human and mouse bona fide stem cells are preferably ...

Generation of Tumor Endothelium Specific Viruses

Disclosed are means, methods, and compositions of matter useful for generation of viruses that selectively grow in the tumor endothelium and causes lysis or augmentation of tumor immunity. In one embodiment an oncolytic virus is selectively maintained in cells resembling tumor endothelium under conditions allowing for the oncolytic virus to capture immunogenic entities found on the cells resembling tumor endothelial cells. In another embodiment, the endothelial cells resembling tumor endothelial cells are utilized as a “Trojan horse” for selective delivery of virus into a tumor or tumor microenvironment.

METHODS FOR PRODUCING CANCER STEM CELL SPHEROIDS

The invention provides a method for producing a population of ready-to-use spheroid forming cancer cells, comprising: (i) growing cancer cells in suspension culture in a first culture medium on one or more first low-adhesion tissue culture plates thereby forming cancer cell spheroids enriched in cancer stem cells; (ii) disaggregating said cancer cell spheroids to form a suspension of single cells enriched in cancer stem cells; (iii) plating said suspension of single cells in a second culture medium on one or more second low-adhesion tissue culture plates; and (iv) freezing said suspension of single cells in said one or more second tissue culture plates, thereby producing a population of ready-to-use spheroid forming cancer cells. Also provided are cell populations produced by the method and kits for growing cancer cell spheroids, including for use in screening of test compound. 1. A method for producing a population of ready-to-use spheroid forming cancer cells , comprising:(i) growing cancer cells in suspension culture in a first culture medium on one or more first low-adhesion tissue culture plates thereby forming cancer cell spheroids enriched in cancer stem cells;(ii) disaggregating said cancer cell spheroids to form a suspension of single cells enriched in cancer stem cells;(iii) plating said suspension of single cells in a second culture medium on one or more second low-adhesion tissue culture plates; and(iv) freezing said suspension of single cells in said one or more second tissue culture plates, thereby producing a population of ready-to-use spheroid forming cancer cells.2. The method according to claim 1 , wherein said cancer cells comprise cells of a cancer cell line or primary cell culture derived from a tumour.3. The method according to claim 2 , wherein the cancer cell line is selected from the group consisting of: human breast carcinoma MDA-MB-436; human glioblastoma U87MG; human colon carcinoma HCT116; human ovarian SK-OV-3; human lung NCI-H446; ...

METHODS FOR DETECTION AND ERADICATION OF MYELOID LEUKEMIA STEM CELLS

There are provided, inter alia, methods and compositions for diagnosis and treatment of acute myeloid leukemia (AML), secondary acute myeloid leukemia (sAML), and age-related diseases. 1. A method for treating acute myeloid leukemia in a subject in need thereof , the method comprising administering to the subject an effective amount of a splicing modulator , thereby treating the acute myeloid leukemia.2. A method for modulating acute myeloid leukemia stem cells , the method comprising contacting the acute myeloid leukemia stem cells with an effective amount of a splicing modulator , thereby modulating the acute myeloid leukemia stem cells.3. The method of claim 2 , wherein normal progenitor cells are not substantially modulated.4. The method of or claim 2 , wherein the method is in vitro or in vivo.5. A method of detecting a protein level in a subject having acute myeloid leukemia claim 2 , the method comprising (i) obtaining a biological sample from the subject; (ii) contacting the biological sample with a detection agent capable of binding at least one protein encoded by at least one RNA set forth in Table 13A and/or Table 13B claim 2 , thereby forming a detectable complex; (iii) detecting and quantitating the detectable complex; and (iv) comparing to a standard control claim 2 , thereby detecting the protein level of the protein in the subject.6. The method of claim 5 , wherein the protein is PTK2B claim 5 , CD44 claim 5 , or a combination thereof.7. The method of claim 5 , further comprising detecting additional protein levels for a plurality of additional proteins encoded by RNA set forth in Table 13A and/or Table 13B by further contacting the biological sample with a plurality of additional different detection agents claim 5 , each additional different detection agent capable of binding to one of the plurality of additional proteins to form a plurality of additional different detectable complexes; and further detecting and quantitating the plurality of ...

BINDING PROTEINS SPECIFIC FOR LOX1 AND USES THEREOF

This disclosure provides LOX1 (LOX1) binding proteins such as anti-LOX1 antibodies, and compositions and methods for making these binding proteins. In certain aspects the LOX1-binding proteins provided herein, inhibit, or antagonize LOX1 activity. In addition, the disclosure provides compositions and methods for diagnosing and treating conditions associated with atherosclerosis, thrombosis, coronary artery disease (CAD), ischemia (e.g., myocardial ischemia), infarction (e.g., myocardial infarction), acute coronary syndrome (ACS), stroke, reperfusion injury, restenosis, peripheral vascular disease, hypertension, heart failure, inflammation (e.g., chronic inflammation), angiogenesis, preeclampsia, cancer and other LOX1-mediated diseases and conditions. 1. (canceled)2. An isolated LOX1-binding protein comprising a set of complementary determining regions (CDRs): heavy chain variable region (VH)-CDR1 , VH-CDR2 , VH-CDR3 , and light chain variable region (VL)-CDR1 , VL-CDR2 and VL-CDR3 , wherein: (ii) VH-CDR2 has the amino acid sequence of SEQ ID NO:2;', '(iii) VH-CDR3 has the amino acid sequence of SEQ ID NO:3;', '(iv) VL-CDR1 has the amino acid sequence of SEQ ID NO:30;', '(v) VL-CDR2 has the amino acid sequence of SEQ ID NO:31; and', '(vi) VL-CDR3 has the amino acid sequence of SEQ ID NO:32;, '(a) (i) VH-CDR1 has the amino acid sequence of SEQ ID NO:1;'} (ii) VH-CDR2 has the amino acid sequence of SEQ ID NO:5;', '(iii) VH-CDR3 has the amino acid sequence of SEQ ID NO:14;', '(iv) VL-CDR1 has the amino acid sequence of SEQ ID NO:30;', '(v) VL-CDR2 has the amino acid sequence of SEQ ID NO:31; and', '(vi) VL-CDR3 has the amino acid sequence of SEQ ID NO:32;, '(b) (i) VH-CDR1 has the amino acid sequence of SEQ ID NO:1;'} (ii) VH-CDR2 has the amino acid sequence of SEQ ID NO:39;', '(iii) VH-CDR3 has the amino acid sequence of SEQ ID NO:44;', '(iv) VL-CDR1 has the amino acid sequence of SEQ ID NO:55;', '(v) VL-CDR2 has the amino acid sequence of SEQ ID NO:60; and', '(vi) VL- ...

METHODS FOR CANCER STEM CELL (CSC) EXPANSION

The invention relates to the methods to increase populations of cancer stem cells (CSCs), including human CSCs, using, for example, a FiSS™ (fiber-inspired smart scaffold) platform, a scaffold for cell culture comprising an electrospun mixture of poly(lactic-co-glycolic acid) (PLGA) and a block copolymer of polylactic acid (PLA) and monomethoxypolyethylene glycol (mPEG). As an example, we demonstrated that MCF-7 cells grown on FiSScsc developed into well-formed single-cell tumoroids (SCTs), showing a ˜3-fold increase in the cancer stem cell (CSC) population versus similar-passage cells grown as monolayers. This increase was further potentiated when the first-generation tumoroids were used to grow second- and third-generation tumoroids. Additionally, we scaled-up the cell culturing protocol from, for example, a 96-well plate to, for example, a 6-well plate, with no loss in the induction of CSCs. We also sorted and froze CSC-enriched cells and successfully thawed them again to grow tumoroids, while maintaining the CSC population. 1. A method for expanding cancer stem cells (CSCs) comprising:a) growing tumoroids on a three-dimensional scaffold in an in vitro cell culture; and,b) isolating CSCs from said tumoroids.2. A method for expanding cancer stem cells (CSCs) comprising:a) growing cancer cells in an in vitro cell culture comprising a three-dimensional scaffold;b) growing tumoroids from said cancer cells on said scaffold;c) harvesting cancer cells from said tumoroids (tumoroid cancer cells);d) transferring said tumoroid cancer cells to a new in vitro cell culture comprising a three-dimensional scaffold;e) growing a subsequent generation of tumoroids from said tumoroid cancer cells on said scaffold of said new in vitro cell culture.3. The method of claim 2 , wherein steps c) through e) are repeated at least once.4. The method of claim 2 , wherein said steps c) through e) are repeated at least twice claim 2 , at least three times claim 2 , at least four times claim 2 ...

Tumor-initiating cells and methods for using same

Isolated and enriched tumor-initiating cell populations, methods for preparing the same, and uses thereof.

METHODS FOR PRODUCING HEPATOCYTES

Methods for producing hepatocytes from pluripotent human stem cells are disclosed herein. The stem cells are plated on a cell culture substrate comprising two laminins. The stem cells are then exposed to different cell culture mediums to induce differentiation. The resulting hepatocytes have higher metabolic capacity compared to hepatocytes cultured on different substrates. 1. A method for producing hepatocytes , comprising:plating pluripotent human stem cells on a cell culture substrate comprising (i) a first laminin which is laminin-521 and (ii) a second laminin selected from the group consisting of laminin-111 and laminin-221, wherein the laminin-521 and the second laminin are each either an intact protein or a protein fragment; andculturing the pluripotent human stem cells on the cell culture substrate to obtain the hepatocytes, wherein the culturing is performed by:culturing the cells in an endoderm differentiation medium containing activin A and Wnt3a;culturing the cells in a hepatic specification medium;culturing the cells in a hepatic differentiation medium; andculturing the cells in a hepatic maturation medium containing hepatocyte growth factor (HGF).2. The method of claim 1 , wherein the weight ratio of the laminin-521 to the second laminin is from about 1:4 to about 1:1.3. The method of claim 1 , wherein the cells are cultured in the endoderm differentiation medium for a period of about 60 hours to about 84 hours.4. The method of claim 1 , wherein the cells are cultured in the hepatic specification medium for a period of about 84 hours to about 108 hours.5. The method of claim 1 , wherein the cells are cultured in the hepatic differentiation medium for a period of about 108 hours to about 132 hours.6. The method of claim 1 , wherein the cells are cultured in the hepatocyte maturation medium for a period of about 84 hours to about 108 hours.7. The method of claim 1 , wherein the activin A is present in the endoderm differentiation medium in an amount of ...

Methods for testing t cell priming efficacy in a subject

The present invention relates to methods for testing T cell priming efficacy in a subject. In particular the present invention relates to an in vitro method for testing T cell priming efficacy in a subject comprising the steps of a) providing sample from the subject, b) culturing the sample in a medium which induces the differentiation of dendritic cells, c) maturing the dendritic cells obtained at step a) in presence of an amount of at least one antigen and an amount of at least one cytokine or ligand suitable for the activation of a pathogen recognition receptor, d) priming and expanding the T cells present in the sample and e) analyzing the func tionality of the primed T cells.

METHODS AND PRODUCTS FOR TRANSFECTION

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed. 1. A method for reprogramming a cell , comprising:a. contacting a cell with a medium containing albumin, wherein the albumin is treated with an ion-exchange resin or charcoal,b. contacting the cell with one or more synthetic RNA molecules, wherein the one or more synthetic RNA molecules includes at least one RNA molecule encoding Oct4 protein, andc. repeating step (b) at least twice during 5 consecutive days.2. The method of claim 1 , wherein the albumin was treated with a short-chain fatty acid.3. The method of claim 2 , wherein the albumin was brought to a temperature of at least 40 C.4. The method of claim 1 , wherein the one or more synthetic RNA molecules further includes at least one RNA molecule encoding at least one member of the group: Sox2 protein claim 1 , Klf4 protein claim 1 , and c-Myc protein.5. The method of claim 1 , wherein the one or more synthetic RNA molecules further includes at least one RNA molecule encoding Sox2 protein claim 1 , at least one RNA molecule encoding Klf4 protein claim 1 , and at least one RNA molecule encoding c-Myc protein.6. The method of claim 1 , further comprising contacting the cell with at least one member of the group: poly-L-lysine claim 1 , poly-L-ornithine claim 1 , RGD peptide claim 1 , fibronectin claim 1 , vitronectin claim 1 , collagen claim 1 , and laminin claim 1 , or a biologically active fragment claim 1 , functional variant or family-member thereof.7. The method of claim 1 , wherein the one or more synthetic RNA molecules contains at least one member of the group: a pseudouridine residue and a 5-methylcytidine residue.8. The method of claim 7 , wherein the cell is a skin cell claim 7 , and the method further ...

Culture medium for expanding breast epithelial stem cells

The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids.

Delta133P53Beta and Delta133P53Gamma Isoforms Are Biomarkers of Cancer Stem Cells

The present invention is in the field of oncology, and more particularly of cancer stem cells. It relates to a method for producing cancer stem cells based on overexpression of Δ133ρ536 isoform, Δ133ρ53γ isoform, or both Δ133ρ536 and Δ133ρ53γ isoforms; a method for predicting the risk that treatment with a chemotherapeutic anti-cancer agent induces cancer stem cells in a subject suffering from cancer from a cancer sample of said subject, based on detection of an increase in Δ133ρ536 isoform, Δ133ρ53γ isoform, or both Δ133ρ536 and Δ133ρ53γ isoforms following chemotherapeutic anti-cancer treatment; to therapeutic uses of a combination of chemotherapeutic anti-cancer agent and an agent reducing Δ133p536 isoform, Δ133ρ53γ isoform, or both Δ133ρ536 and Δ133ρ53γ isoforms expression; and also to screening methods for anti-cancer stem cells agents. 1. A method for producing cancer stem cells , comprising:a) transducing cancer cells with a vector expressing Δ133p53β isoform, Δ133p53γ isoform, or both Δ133p53β and Δ133p53γ isoforms;b) culturing transduced cancer cells in a medium supporting expansion of transduced cancer cells; andc) isolating cancer stem cells.2. The method according to claim 1 , wherein cancer cells are selected from solid cancer cells claim 1 , preferably from breast cancer cells claim 1 , colorectal cancer cells claim 1 , ovarian cancer cells claim 1 , digestive cancers cells claim 1 , pancreatic cancer cells claim 1 , lung cancer cells claim 1 , prostate cancer cells and throat cancer cells; or from hematopoietic cancer cells claim 1 , preferably from leukaemia cells or lymphoma cells.3. The method according to or claim 1 , wherein the vector expressing Δ133p53β isoform claim 1 , Δ133p53γ isoform claim 1 , or both Δ133p53β and Δ133p53γ isoforms is a retroviral vector claim 1 , preferably a Murine Stem Cell Virus (MSCV) vector claim 1 , comprising a nucleic acid molecule encoding Δ133p53β isoform claim 1 , Δ133p53γ isoform claim 1 , or both Δ133p53β and Δ ...

COMPOSITION FOR INDUCING DEDIFFERENTIATION INTO CANCER STEM CELLS COMPRISING RIBOSOME-ACTIVATING INHIBITOR AS ACTIVE INGREDIENT, CANCER ORGANOID CULTURE METHOD AND ANTICANCER DRUG SCREENING PLATFORM BASED THEREON

A composition for inducing dedifferentiation from cancer cells to cancer stem cells comprising a ribosome-activating inhibitor as an active ingredient, a method of culturing a cancer organoid based thereon and an anticancer drug screening platform, and the increase of colorectal cancer stem cell group induced by the exposure of ribosome-inactivating stress was regulated by the ATF3 gene. 1. A method of inducing dedifferentiation from cancer cells to cancer stem cells comprising: a step of treating cancer cells with a ribosome-activating inhibitor.2. The method of inducing dedifferentiation from cancer cells to cancer stem cells of claim 1 , wherein the ribosome-activating inhibitor is anisomycin (ANS claim 1 , RIS-1) or deoxynivalenol (DON claim 1 , RIS-2).3. The method of inducing dedifferentiation from cancer cells to cancer stem cells of claim 1 , wherein the cancer is colorectal cancer (CRC).4. A method of forming cancer organoid induced by ribosome-inactivating stress comprising:a step of forming spheroids by pretreating cancer cells with a ribosome-activating inhibitor; anda step of forming cancer organoid by culturing formed spheroids.5. The method of forming cancer organoid induced by ribosome-inactivating stress of claim 4 , wherein the cancer is colorectal cancer (CRC).6. The method of forming cancer organoid induced by ribosome-inactivating stress of claim 4 , wherein the ribosome-activating inhibitor is anisomycin (ANS claim 4 , RIS-1) or deoxynivalenol (DON claim 4 , RIS-2).7. A method of screening anticancer drug resistance inhibitor comprising:{'claim-ref': {'@idref': 'CLM-00004', 'claim 4'}, 'a step of contacting cancer organoid induced by ribosome-inactivating stress formed according to the method of with a test substance;'}a step of measuring anticancer drug susceptibility to the cancer organoid induced by ribosome-inactivating stress in contact with the test substance; anda step of selecting a test substance having increased anticancer drug ...

Method for Isolating Cancer Stem Cells

The invention relates to the use of a lectin that recognizes the fucose α 1-2 galactose unit, as a first means for labeling and optionally a second means for labelling colorectal cancer stem cells, in particular a lectin that recognizes the T antigen, in order to carry out a method for the detection and optionally isolation of colorectal cancer stem cells, a method for the detection and optionally isolation of colorectal cancer stem cells for research purposes, and a method for the in vitro diagnosis of colorectal cancer recurrence risk and/or aggressiveness so as to define a prognostic value in order to make colorectal cancer therapy adjustments, as well as a kit comprising a lectin that recognizes the fucose α 1-2 galactose unit and a lectin that recognizes the T antigen. 118-. (canceled)19. A method of detecting or isolating colorectal cancer stem cells in a biological sample comprising colorectal cells , the method comprising the step of contacting the biological sample in vitro with a lectin that binds fucose α 1-2 galactose.20Ulex EuropaeusTrichosanthes Japonica. The method of claim 19 , wherein the lectin that binds fucose α 1-2 galactose is selected from the group consisting of Agglutinin I (UEA-1) and Agglutinin II (TJA-II).21. The method of claim 19 , the method further comprising the step of contacting the biological sample in vitro with means of labeling colorectal cancer stem cells or with at least one lectin that binds to T antigen.22Amaranthus CaudatusAgaricus Bisporus. The method of claim 21 , wherein the lectin that binds to T antigen is selected from the group consisting of Lectin (ACA) claim 21 , Agglutinin (ABA) claim 21 , Jacalin claim 21 , and a mixture of lectins that binds T antigen selected from the group consisting of ABA and ACA claim 21 , ABA and Jacalin claim 21 , and Jacalin and ACA.23. A kit for detecting or isolating colorectal cancer stem cells in a biological sample comprising colorectal cells claim 21 , the kit comprising a lectin ...