Cell culture platform for single cell sorting and enhanced reprogramming of iPSCs
AssigneeFate Therapeutics, Inc.
InventorsBahram Valamehr, Ramzey Abujarour, Peter Flynn
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Abstract
The invention provides cell culture conditions for culturing stem cells, including feeder-free conditions for generating and culturing human induced pluripotent stem cells (iPSCs). More particularly, the invention provides a culture platform that allows long-term culture of pluripotent cells in a feeder-free environment; reprogramming of cells in a feeder-free environment; single-cell dissociation of pluripotent cells; cell sorting of pluripotent cells; maintenance of an undifferentiated status; improved efficiency of reprogramming; and generation of a naïve pluripotent cell.
Claims11
A method of sorting a population of human cells to obtain a population of cells enriched for pluripotent cells comprising:
- a) suspending a mixed population of dissociated cells to thereby produce a suspension comprising the mixed population of cells, wherein the mixed population of cells comprises human induced pluripotent stem cells (hiPSCs) having ground state pluripotency and human non-pluripotent cells;
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- b) sorting the cells in the suspension for cells expressing one or both of SSEA4 and TRA181, thereby obtaining an enriched population of cells enriched for the hiPSCs, wherein the hiPSCs of the enriched population of cells comprise the hiPSCs having ground state pluripotency;
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- wherein the hiPSCs having ground state pluripotency in the mixed population of cells and the enriched population of cells have the ability to reactivate GATA6, CDX2, and CGB upon differentiation.
Dependencies are inferred from the claim text.