Differentiation of pluripotent embryonic stem cells
AssigneeWisconsin Alumni Research Foundation
InventorsJames Thomson, Thomas 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 mesoendoderm cells by treatment with caspase-like inhibitors. Also described is a population of mesoendoderm cells obtained therefrom. The embryonic stem cell derived mesoendoderm cells have the general morphological and cell surface marker characteristics of mesoendoderm cells.
Claims13
A method for directing the differentiation of primate embryonic stem cells into mesoendoderm cells comprising the steps of
- a) culturing a culture of embryonic stem cells in a medium containing an effective amount of at least one caspase inhibitor;
- and
- b) sub-culturing cells which exhibit a morphology of mesoendoderm cells.
A cell culture derived from primate embryonic stem cells, the culture comprising predominantly mesoendoderm cells have the following characteristics:
- uniform spindle shaped-morphology with a much smaller nucleus in comparison to undifferentiated embryonic stem cells, reduced caspase activity and reduced ability to bind Annexin-V.
A cell culture of mesoendoderm cells derived from primate embryonic stem cells, the culture composed of cells over 90% of which test negative for a caspase-3 activity.
A method of selecting for a cell population enriched for pluripotent cells, the method comprising assaying a culture of embryonic stem cells for the presence of a protein marker exhibiting caspase activity and selecting only those cells which have reduced caspase activity.
A method for maintaining embryonic stem cells in a pluripotent culture comprising the steps of:
- a) contacting the cell culture with an amount of an agonist of a apoptotic pathway effective to inhibit differentiation of the cell;
- and
- b) exposing the cell culture to cell growth conditions such that the cells proliferate.
Dependencies are inferred from the claim text.