Methods and compositions for inducing hematopoietic cell differentiation
AssigneeFate Therapeutics, Inc.
InventorsBahram Valamehr, Raedun Clarke
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Abstract
The invention provides culture platforms, cell media, and methods of differentiating pluriptent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein, which enable feed-free, monolayer culturing and in the absence of EB formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, and NK cells.
Claims15
A differentiation platform for generating human pluripotent stem cell-derived T lineage cells comprising:
- (i) a culture medium that comprises a BMP activator, wherein the medium is suitable for differentiating and expanding human pluripotent stem cell-derived mesodermal cells from human pluripotent stem cells;
- (ii) a culture medium comprising a BMP activator, bFGF, and a GSK3 inhibitor, wherein the medium is suitable for obtaining definitive hemogenic endothelium (HE) potential in the human pluripotent stem cell-derived mesodermal cells;
- (iii) a culture medium comprising a ROCK inhibitor and one or more growth factors and cytokines selected from the group consisting of bFGF, VEGF, SCF, IL6, and IL11, wherein the medium is suitable for differentiating and expanding human pluripotent stem cell-derived definitive hemogenic endothelium from the human pluripotent stem cell-derived mesodermal cells with hemogenic endothelium potential;
- (iv) a culture medium comprising a ROCK inhibitor, IL7, and one or more growth factors and cytokines selected from the group consisting of VEGF, bFGF, SCF, and Flt3L, wherein the culture medium is suitable for differentiating human pluripotent stem cell-derived definitive hemogenic endothelium into human pluripotent stem cell-derived pre-T cell progenitors;
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- (v) a culture medium comprising one or more growth factors and cytokines selected from the group consisting of SCF, Flt3L, and IL7, wherein the culture medium is free of one or more of VEGF, bFGF, BMP activators and ROCK inhibitors, wherein the culture medium is suitable for differentiating human pluripotent stem cell-derived pre-T cell progenitors to human pluripotent stem cell-derived T cell progenitors or T cells.
Dependencies are inferred from the claim text.