Method for making hematopoietic cells
First Claim
1. A method for differentiating human pluripotent stem (hPS) cells into a cell population with hematopoietic potential, comprising:
- a) harvesting undifferentiated hPS cells from a feeder-free culture;
b) differentiating the harvested hPS cells in a culture environment essentially free of any cells having a different genotype, but containing at least two hematopoietic growth factors selected from stem cell factor (SCF), FLT-3 ligand, IL-3, IL-6, and granulocyte colony stimulating factor (G-CSF);
c) simultaneously or subsequently culturing the differentiating cells with a bone morphogenic protein; and
d) harvesting from the culture environment a cell population that is at least 1% CD45 positive, or that forms colonies in an assay for hematopoietic colony forming units (CFU) at a plating efficiency of at least −
1 in 2000.
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Abstract
This invention provides a system for producing cells of the hematopoietic lineage from embryonic stem cells. Differentiation is conducted in the presence of hematogenic cytokines and other factors listed in the disclosure. The cell population that is obtained is remarkably enriched in CD45 +ve cells, a marker of early hematopoietic precursor with self-renewing capacity. Including a bone morphogenic protein during the differentiation process enhances the ability of the cell population to form secondary colonies. Because of the enormous replicative capacity of embryonic stem cells, this provides an important new commercial source of hematopoietic cells.
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Citations
15 Claims
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1. A method for differentiating human pluripotent stem (hPS) cells into a cell population with hematopoietic potential, comprising:
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a) harvesting undifferentiated hPS cells from a feeder-free culture;
b) differentiating the harvested hPS cells in a culture environment essentially free of any cells having a different genotype, but containing at least two hematopoietic growth factors selected from stem cell factor (SCF), FLT-3 ligand, IL-3, IL-6, and granulocyte colony stimulating factor (G-CSF);
c) simultaneously or subsequently culturing the differentiating cells with a bone morphogenic protein; and
d) harvesting from the culture environment a cell population that is at least 1% CD45 positive, or that forms colonies in an assay for hematopoietic colony forming units (CFU) at a plating efficiency of at least −
1 in 2000. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification