Methods and compositions for inducing hematopoietic cell differentiation
AssigneeFate Therapeutics, Inc.
InventorsBahram VALAMEHR, Raedun CLARKE
Life of this patent
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Abstract
The invention provides culture platforms, cell media, and methods of differentiating pluripotent 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.
Claims21
-98. (canceled)
An in vitro method for generating human pluripotent stem cell-derived definitive hemogenic endothelium, the method comprising:
- (i) contacting human pluripotent stem cells with a composition comprising a BMP activator, and optionally bFGF, to allow cell differentiation and expansion to obtain human pluripotent stem cell-derived mesodermal cells;
- (ii) contacting the human pluripotent stem cell-derived mesodermal cells with a composition comprising a BMP activator, bFGF, and a GSK3 inhibitor, to allow cell differentiation and expansion to obtain human pluripotent stem cell-derived mesodermal cells having definitive hemogenic endothelium (HE) potential, wherein the composition is optionally free of TGFβ receptor/ALK inhibitor;
- and
- (iii) contacting the human pluripotent stem cell-derived mesodermal cells having definitive HE potential with a composition comprising a ROCK inhibitor, and one or more growth factors and cytokines selected from the group consisting of bFGF, VEGF, SCF, IL6 and IL11, to allow cell differentiation and expansion to obtain human pluripotent stem cell-derived definitive hemogenic endothelium, wherein the composition is optionally free of TGFβ receptor/ALK inhibitor.
A differentiation platform for generating human pluripotent stem cell-derived definitive hemogenic endothelium, the differentiation platform 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;
- and
- (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 definitive HE potential.
Dependencies are inferred from the claim text.