Differentiation of Multi-Lineage Progenitor Cells to Pancreatic Cells
First Claim
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1. A method of producing a population of cells having a pancreatic cell phenotype, said method comprising:
- a) providing a collagen-coated culturing device housing a purified population of human fetal blood multi-lineage progenitor cells (MLPC) or a clonal line of MLPC, wherein said MLPC are positive for CD9, CD13, CD29, CD44, CD73, CD90, and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3, and SSEA-4;
b) culturing said purified population of MLPC or said clonal line of MLPC with a differentiation medium until cells having a hepatocytye-pancreatic precursor phenotype are obtained, said differentiation medium comprising ascorbic acid, hydrocortisone, transferrin, insulin, epidermal growth factor, hepatocyte growth factor, and stem cell factor; and
c) further culturing said cells having said hepatocytye-pancreatic precursor phenotype in said differentiation medium in the presence of retinoic acid to obtain cells having said pancreatic cell phenotype,.
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Abstract
Fetal blood multi-lineage progenitor cells that are capable of a wide spectrum of transdifferentiation are described, as well as methods of differentiating the progenitor cells into pancreatic cells.
105 Citations
21 Claims
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1. A method of producing a population of cells having a pancreatic cell phenotype, said method comprising:
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a) providing a collagen-coated culturing device housing a purified population of human fetal blood multi-lineage progenitor cells (MLPC) or a clonal line of MLPC, wherein said MLPC are positive for CD9, CD13, CD29, CD44, CD73, CD90, and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3, and SSEA-4; b) culturing said purified population of MLPC or said clonal line of MLPC with a differentiation medium until cells having a hepatocytye-pancreatic precursor phenotype are obtained, said differentiation medium comprising ascorbic acid, hydrocortisone, transferrin, insulin, epidermal growth factor, hepatocyte growth factor, and stem cell factor; and c) further culturing said cells having said hepatocytye-pancreatic precursor phenotype in said differentiation medium in the presence of retinoic acid to obtain cells having said pancreatic cell phenotype,. - View Dependent Claims (2, 3)
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4. A method for producing a population of cells having a pancreatic cell phenotype from human fetal blood, said method comprising:
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a) contacting a human fetal blood sample with a composition, said composition comprising; i) dextran; ii) anti-glycophorin A antibody; iii) anti-CD15 antibody; and iv) anti-CD9 antibody; b) allowing said sample to partition into an agglutinate and a supernatant phase; c) recovering cells from said supernatant phase; d) purifying MLPC from the recovered cells by adherence to a solid substrate, wherein said MLPC are positive for CD9 and positive for CD45; e) culturing said MLPC such that said MLPC obtain a fibroblast morphology, wherein said MLPC having said fibroblast morphology are positive for CD9, CD13, CD29, CD44, CD73, CD90, and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3, and SSEA-4; f) culturing said MLPC having said fibroblast morphology with a differentiation medium effective to induce differentiation of said MLPC into cells having a hepatocyte-pancreatic precursor cell phenotype, said differentiation medium comprising ascorbic acid, hydrocortisone, transferrin, insulin, epidermal growth factor, hepatocyte growth factor, and stem cell factor; and g) culturing said cells having a hepatocyte-pancreatic precursor cell phenotype in said differentiation medium in the presence of retinoic acid to obtain cells having said pancreatic cell phenotype. - View Dependent Claims (5, 6)
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- 7. A clonal population of cells having a pancreatic cell phenotype.
- 19. A composition comprising a purified population of MLPC or a clonal line of human fetal blood MLPC and a differentiation medium effective to induce differentiation of said MLPC into cells having a pancreatic cell phenotype, wherein said MLPC are positive for CD9, CD13, CD29, CD44, CD73, CD90, and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3, and SSEA-4.
Specification