Differentiation of pluripotent embryonic stem cells
AssigneeWisconsin Alumni Research Foundation
InventorsJames A. Thomson, Thomas P. Zwaka
Life of this patent
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Abstract
The invention relates to a method to induce primate embryonic stem cells to differentiate into a relatively homogenous population of mesendoderm cells by treatment with caspase-like inhibitors. Also described is a population of mesendoderm cells obtained therefrom. The embryonic stem cell derived mesendoderm cells have the general morphological and cell surface marker characteristics of mesendoderm cells.
Claims12
A method for directing the differentiation of human embryonic stem cells into a population of multipotent cells comprising the steps of
- culturing a population of human embryonic stem cells for 2 to 3 days in a culture medium containing at least one caspase inhibitor that inhibits cleavage of poly(ADPribosyl) polymerase (PARP-1), wherein the inhibitor concentration is between 80 μM to 200 μM;
- and
- yielding the population of multipotent cells.
A multipotent cell population, wherein at least 75% of the cells are Oct4 negative, and exhibit uniform, spindle shaped-morphology with a small nucleus, reduced caspase activity, reduced ability to bind Annexin-V, and increased expression of mesoderm and endoderm genes selected from the group consisting of brachyury variant A, BMP4, GATA-3, WNT5A, WNT3, Nodal, Scl, Flk and DKK4 relative to a population of human embryonic stem cells.
A method of producing a population of multipotent cells comprising the steps of
- a) providing a culture of human embryonic stem cells;
- b) culturing the cell population of step (a) for 2 to 3 days in a culture medium containing at least one caspase inhibitor that inhibits cleavage of poly(ADPribosyl) polymerase PARP-1), and inhibits caspase-3 activity, wherein the inhibitor concentration is between 80 μM to 200 μM;
- and
- (c) yielding the population of multipotent cells.
Dependencies are inferred from the claim text.