METHODS AND DEVICES FOR ISOLATING EMBRYONIC STEM CELLS

Methods, devices and kits are provided for isolating or enriching multicellular embryonic stem (ES) cell colonies from a mixture of multicellular ES cell colonies and single ES cells present in a cellular suspension, by utilizing a filtration matrix that selectively excludes passage of multicellular ES cell colonies. Isolated or enriched multicellular ES cell colonies can be used for propagating pluripotent ES cells. 1. A method for isolating or enriching multicellular ES cell colonies from a mixture of multicellular ES cell colonies and single ES cells present in a cellular suspension comprising the steps of:a. providing a filtration matrix through which single ES cells but not multicellular ES cell colonies can pass from a first side to a second side;b. contacting the cellular suspension with the first side of the filtration matrix;c. passing the cellular suspension from the first side to the second side of the filtration matrix; andd. collecting the multicellular ES cell colonies from the first side of the filtration matrix, thereby isolating or enriching multicellular ES cell colonies therefrom.2. The method of wherein the filtration matrix comprises polyester claim 1 , polyamide claim 1 , aramid claim 1 , acrylic claim 1 , or PTFE.3. The method of wherein the filtration matrix has a porosity of about 20 microns to about 30 microns.4. The method of wherein the passing is via centrifugation claim 1 , application of positive pressure to the first side of the filtration matrix claim 1 , application of negative pressure to the second side of the filtration matrix claim 1 , or gravity filtration.5. The method of wherein the collecting comprises the steps of:a. discarding the single ES cells from the second side of the filtration matrix;b. passing a liquid medium from the second side to the first side of the filtration matrix, whereby multicellular ES cell colonies retained on the first side are suspended in the liquid medium; andc. collecting the liquid medium.6. The ...

METHODS FOR IDENTIFYING, ISOLATING, AND UTILIZING ENDOCRINE PROGENITOR CELLS FROM ADULT HUMAN PANCREAS

The presence of the cell surface marker CD133 or the presence of a glycosylated form of the prominin- gene product on adult pancreatic cells is used to identify pancreatic endocrine progenitor cells, and useful in methods of isolation and enrichment. Isolated pancreatic endocrine progenitor cells can be used for cell based therapy for insulin-dependent diabetes and pancreatectomy patients. 1. A method for identifying pancreatic endocrine progenitor cells in a cellular sample or tissue sample comprising exposing the cellular sample or tissue sample to at least one detectable agent that binds to cells expressing a glycosylated form of a prominin-1 gene product , thereby identifying pancreatic endocrine progenitor cells therein.2. The method of wherein the at least one detectable agent is a detectably labeled primary binding partner that binds to a glycosylated form of a prominin-1 gene product claim 1 , or a combination of an unlabeled primary binding partner that binds to a glycosylated form of a prominin-1 gene product and a detectably labeled secondary binding partner that binds to the unlabeled primary binding partner.3. The method of wherein the at least one detectable agent comprises at least one antibody or lectin that binds to a glycosylated form of a prominin-1 gene product claim 1 , binds to CD133 claim 1 , binds to epitope CD133/1 claim 1 , or binds to epitope CD133/2.4. The method of wherein the cellular sample is a pancreatic islet preparation.5. The method of wherein the cellular sample is human pancreatic tissue.6. A method for isolating pancreatic endocrine progenitor cells from a cellular population comprising identifying cells expressing a glycosylated form of a prominin-1 gene product therein and separating said cells from the cellular population claim 1 , thereby isolating pancreatic endocrine progenitor cells.7. The method of wherein the identifying comprises exposing the cellular population to at least one detectable agent that labels cells that ...

COMPOSITIONS AND METHODS FOR GROWING HUMAN EMBRYONIC CELLS

Methods for deriving and cultivating human embryonic stem (ES) cells and maintaining their pluripotency in culture is provided by utilizing secreted products obtained from the culture medium of human embryonic gem (EG) cell derivatives, such as embryoid body-derived cells. Substrates include compounds such as collagen I, fibronectin, or superfibronectin, or extracellular matrix, typically human derived. 1. A method for cultivating human embryonic stem (ES) cells and maintaining the pluripotency thereof comprising growing the human embryonic stem (ES) cells in a culture medium comprising secreted products from human embryonic germ (EG) cell derivatives.2. The method of wherein the human embryonic germ (EG) cell derivatives are embryoid body-derived cells.3. The method of wherein said human embryoid body-derived cells are LVEC cells or SDEC cells.4. The method of further comprising a substrate.5. The method of wherein the substrate is collagen I claim 4 , collagen IV claim 4 , fibronectin claim 4 , superfibronectin claim 4 , laminin claim 4 , heparan sulfate proteoglycan claim 4 , entactin claim 4 , or any combination thereof.6. The method of wherein the collagen I is human type 1 collagen.7. The method of wherein the substrate comprises a synthetic or biosynthetic cell adhesion molecule or a mixture thereof.8. The method of further comprising an extracellular matrix.9. The method of wherein the extracellular matrix is obtained from human embryonic germ (EG) cell derivatives.10. The method of wherein the human embryonic germ (EG) cell derivatives are human embryoid body-derived cells.11. The method of wherein the human embryoid body-derived cells are LVEC cells or SDEC cells.12. The method of wherein the extracellular matrix is EHS mouse sarcoma basement membrane or human extracellular matrix.13. The method of wherein the human extracellular matrix is obtained from human mesenchymal stem cells claim 12 , cells derived from human umbilical cord blood or human ...

MULTI-INDICATION MRNA CANCER IMMUNOTHERAPY

Synthetic bacterial messenger RNA can be used to prepare autologous, allogenic or direct nucleic acid cancer vaccines. Cancer cells are transfected either in vitro or in vivo with mRNA obtained from DNA that encodes an immunogenic bacterial protein. An immune response to the cancer is generated from direct administration of the mRNA in vivo or administration of vaccines prepared from cancer cells in vitro. 1. A method of treating a cancer patient , comprising administering a ribonucleic acid having the sequence of SEQ ID NO: 13 , SEQ ID NO: 15 or SEQ ID NO: 16 directly into a tumor or tumor draining lymph node of a cancer patient or administering a composition comprising tumor cells transformed with said nucleic acid sequences.2. The method of wherein the cancer patient tumor is a carcinoma claim 1 , sarcoma claim 1 , myeloma claim 1 , lymphoma claim 1 , or leukemia.3. The method of wherein the transformed tumor cells are carcinoma claim 1 , sarcoma claim 1 , myeloma claim 1 , lymphoma claim 1 , leukemia or mixtures of said cells.4. The method of wherein at least one of the transformed tumor cells is the same as the tumor of the cancer patient.5. The method of wherein direct administration of said ribonucleic acid is by needle or needleless injection.6. The method of wherein administration of the composition comprising said transformed tumor cells is by intradermal claim 1 , subcutaneous claim 1 , intravenous or intranodal administration.7. The method of wherein the cancer patient is treated prior to or concurrently with additional therapy selected from the group consisting of chemotherapy claim 1 , radiation therapy claim 1 , checkpoint inhibitor therapy claim 1 , whole cell vaccine claim 1 , nucleic acid therapy other than a nucleic acid having the sequence of SEQ ID NO: 13 claim 1 , SEQ ID NO: 15 or SEQ ID NO: 16 claim 1 , natural killer cell therapy claim 1 , and chimeric antigen receptor therapy.8. The method of wherein the cancer patient is treated prior to or ...

HUMAN ENDOCRINE PROGENITORS FROM ADULT PANCREATIC TISSUE

The present invention relates to the field of progenitor cells. More specifically, the present invention provides compositions and methods for isolating endocrine progenitor cells from pancreatic tissue. In certain embodiments, the method comprises the steps of (a) providing a pancreatic tissue sample; (b) isolating cells positive for CD133+; and (c) culturing the isolated cells in defined media for at least about 4 days. 1. A method for isolating a population of endocrine progenitor cells comprising the steps of:a. providing a pancreatic tissue sample;b. isolating cells positive for CD133+; andc. culturing the isolated cells in defined media for at least about 4 days.2. The method of claim 1 , wherein the isolation step is carried out using immunomagnetic beads.3. The method of claim 1 , wherein the isolation step is accomplished using fluorescence-activate cell sorting (FACS).4. The method of claim 3 , further comprising selecting for Aldefluor-positive cells prior to the culturing step.5. The method of claim 1 , further comprising selecting for SSEA-4+ cells prior to the culturing step.6. A substantially pure population of CD133+ cells isolated by the method of claim 1 , wherein the cells are also NGN3+.7. A population of cells comprising at least about 90% endocrine progenitor cells claim 1 , wherein the progenitor cells have the phenotype CD133+.8. The population of cells of claim 7 , wherein the progenitor cells have the phenotype NGN3+.9. The population of cells of claim 7 , wherein the progenitor cells are ALDH+.10. The population of cells of claim 7 , wherein the progenitor cells are SSEA-4+.11. The population of cells of claim 7 , wherein the progenitor cells are capable of clonal pancosphere formation.12. A method for differentiating human endocrine progenitors comprising the steps of:a. suspending CD133+/NGN3+ cells in a matrix;b. mixing the cells-matrix with in a fiber mesh; andc. differentiating the cells into CPEP+ cells.13. The method of claim 12 , ...

MODIFIED MRNA FOR MULTICELL TRANSFORMATION

Synthetic bacterial messenger RNA can be used to prepare autologous, allogenic or direct nucleic acid cancer vaccines. Cancer cells are transfected either in vitro or in vivo with mRNA obtained from DNA that encodes an immunogenic bacterial protein. An immune response to the cancer is generated from direct administration of the mRNA in vivo or administration of vaccines prepared from cancer cells in vitro. Codon modification of the mRNA can optimize expression of an immunogenic polypeptide in cancer cells. 1. A codon optimized ribonucleic acid for expression in human cells that expresses the polynucleotide encoded by SEQ ID NO: 14 in a cell transformed with the ribonucleic acid.2. The codon optimized ribonucleic acid of which is selected from the group consisting of SEQ ID NO: 17 claim 1 , SEQ ID NO: 18 and SEQ ID NO: 19.3. The codon optimized ribonucleic acid of wherein the transformed cell is a cancer cell.4. The codon optimized ribonucleic acid of wherein the transformed cancer cell induces an immunogenic response when introduced into a cancer patient.5. The codon optimized ribonucleic acid of wherein the cancer cell is selected from a carcinoma claim 3 , sarcoma claim 3 , myeloma claim 3 , or lymphoma cell or mixtures of two or more of said cells.6. The codon optimized ribonucleic acid of which expresses an immunogenic polynucleotide when introduced into a tumor or tumor draining lymph node in vivo.7. The codon optimized ribonucleic acid of wherein the tumor or tumor draining lymph node comprises carcinoma claim 6 , sarcoma claim 6 , myeloma claim 6 , or lymphoma cells.8. A synthetic ribonucleic acid comprising a proximal design element which increases expression of a polypeptide having the sequence of SEQ ID NO: 14 in a mammalian cell transformed with the ribonucleic acid.9. The synthetic ribonucleic acid of wherein the proximal design element is the translation initiation sequence.10. The synthetic ribonucleic acid of selected from the group consisting of SEQ ...

Human embryoid body-derived cells

Compositions and methods for growing human embryonic stem cells

Methods for deriving and cultivating human embryonic stem (ES) cells and maintaining their pluripotency in culture is provided by utilizing secreted products obtained from the culture medium of human embryonic germ (EG) cell derivatives, such as embryoid body-derived cells. Substrates include compounds such as collagen I, fibronectin, or superfibronectin, or extracellular matrix, typically human derived.

Multi-indication mRNA cancer immunotherapy

Synthetic bacterial messenger RNA can be used to prepare autologous, allogenic or direct nucleic acid cancer vaccines. Cancer cells are transfected either in vitro or in vivo with mRNA obtained from DNA that encodes an immunogenic bacterial protein. An immune response to the cancer is generated from direct administration of the mRNA in vivo or administration of vaccines prepared from cancer cells in vitro.

Methods for identifying, isolating, and utilizing endocrine progenitor cells from adult human pancreata

The presence of the cell surface marker CD 133 or the presence of a glycosylated form of the prominin- 1 gene product on adult pancreatic cells is used to identify pancreatic endocrine progenitor cells, and useful in methods of isolation and enrichment. Isolated pancreatic endocrine progenitor cells can be used for cell based therapy for insulin-dependent diabetes and pancreatectomy patients.

Modified mRNA for multicell transformation

Synthetic bacterial messenger RNA can be used to prepare autologous, allogenic or direct nucleic acid cancer vaccines. Cancer cells are transfected either in vitro or in vivo with mRNA obtained from DNA that encodes an immunogenic bacterial protein. An immune response to the cancer is generated from direct administration of the mRNA in vivo or administration of vaccines prepared from cancer cells in vitro. Codon modification of the mRNA can optimize expression of an immunogenic polypeptide in cancer cells.

Multi-indication mrna cancer immunotherapy

Methods for identifying, isolating, and utilizing endocrine progenitor cells from adult human pancreata

The presence of the cell surface marker CD133 or the presence of a glycosylated form of the prominin-1 gene product on adult pancreatic cells is used to identify pancreatic endocrine progenitor cells, and useful in methods of isolation and enrichment. Isolated pancreatic endocrine progenitor cells can be used for cell based therapy for insulin-dependent diabetes and pancreatectomy patients.

Exosome delivery of cancer therapeutics

Chimeric expression constructs encoding an immunogenic polypeptide fused to a lactadherin domain are described. The compositions are useful for delivery of antigens to different cells, including cancer cells.

Modified mrna for multicell transformation

Synthetic bacterial messenger RNA can be used to prepare autologous, allogenic or direct nucleic acid cancer vaccines. Cancer cells are transfected either in vitro or in vivo with mRNA obtained from DNA that encodes an immunogenic bacterial protein. An immune response to the cancer is generated from direct administration of the mRNA in vivo or administration of vaccines prepared from cancer cells in vitro. Codon modification of the mRNA can optimize expression of an immunogenic polypeptide in cancer cells.

Modified mrna for multicell transformation