Methods and compositions for inducing hematopoietic cell differentiation
AssigneeFate Therapeutics, Inc.
InventorsBahram Valamehr, Raedun Clarke, Ryan Bjordahl
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, NK cells, NKT cells and B cells.
Claims31
1-51. (canceled)
A method of identifying definitive hemogenic endothelium (HE) cells in pluripotent stem cell differentiation, comprising:
- (i) obtaining a cell population undergoing differentiation;
- (ii) introducing to the cell population an antibody specific to CD93;
- and
- (iii) identifying a subpopulation of cells in the cell population having CD93 expression level less than 1%, wherein the subpopulation of cells comprise definitive hemogenic endothelium cells.
A method of generating pluripotent stem cell derived definitive hemogenic endothelium (HE) cells, comprising culturing pluripotent stem cell derived mesodermal cells having definitive hemogenic endothelium (HE) potential in a medium comprising a Wnt pathway agonist to obtain definitive HE cells, wherein the obtained definitive HE cells:
- (i) are increased in numbers and percentage in cell population;
- (ii) have increased potency in differentiation;
- and/or
- (iii) improved HE cellularity,
- as compared to culturing without the Wnt pathway activator.
A method of generating antigen-specific induced pluripotent stem cells (iPSC) or derivative hematopoietic lineage cells therefrom comprising:
- a) isolating antigen specific T cells from a selected source that is donor-, disease- or treatment response-specific;
- b) reprogramming the antigen-specific T cells to obtain pluripotent stem cells;
- and
- c) directing differentiation of the pluripotent stem cells of step b) to hematopoietic lineage cells, wherein directing differentiation comprises:
- (i) contacting the pluripotent stem cells with a composition comprising a BMP pathway activator, and optionally bFGF, to obtain mesodermal cells;
- and
- (ii) contacting the mesodermal cells with a composition comprising a BMP pathway activator, bFGF, and a WNT pathway activator, to obtain mesodermal cells having definitive hemogenic endothelium (HE) potential;
- wherein mesodermal cells and mesodermal cells having definitive HE potential are obtained in steps (i) and (ii) without the step of forming embryoid bodies.
81-141. (canceled)
Dependencies are inferred from the claim text.