Induction of Hemogenic Endothelium from Pluripotent Stem Cells
AssigneeWisconsin Alumni Research Foundation
InventorsIgor Slukvin, Irina Elcheva
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Abstract
Described herein are methods and related compositions for inducing differentiation of human pluripotent stem cells (hPSCs) into hemogenic endothelium with pan-myeloid potential or restricted potential, by forced expression in the hPSCs of a combination of transcription factors as described herein.
Claims22
A method for generating human hemogenic endothelial cells with pan-myeloid potential, comprising
- (i) forcing expression, in human pluripotent stem cells, of:
- (a) an ETV2 or ERG protein, and a GATA1 protein, or functional homologs thereof;
- (b) an ETV2 or ERG protein, and a GATA2 protein, or functional homologs thereof;
- or
- (c) an ETV2 or ERG protein, and a GFI1 protein, or functional homologs thereof;
- and
- (ii) culturing the human pluripotent stem cells following step (i), under culture conditions that support expansion of hematopoietic cells, to obtain hemogenic endothelial cells that are VE-cadherin+, CD226+, and CD73−.
A method for generating human hemogenic endothelial cells with restricted erythroid, megakaryocytic, and macrophage potential, comprising
- (i) forcing expression, in human pluripotent stem cells, of a TAL1 protein, and a GATA2 protein or GATA1 protein or functional homologs thereof;
- and
- (ii) culturing the human pluripotent stem cells following step (i), under culture conditions that support expansion of hematopoietic cells, to obtain hemogenic endothelial cells that are VE-cadherin+, CD226+, CD73−, and have restricted erythroid, megakaryocytic, and macrophage potential.
A recombinant human pluripotent stem cell comprising:
- (i) one or more exogenous nucleic acids suitable for expression of
- (a) an ETV2 or ERG protein, and a GATA1 protein, or functional homologs thereof;
- (b) an ETV2 or ERG protein, and a GATA2 protein, or functional homologs thereof;
- or (c) an ETV2 or ERG protein, and a GFI1 protein
- (ii) exogenous polypeptides comprising the amino acid sequences of any of (a), (b), or (c).
A kit for hemogenic reprogramming, comprising:
- (i) one or more isolated nucleic acids comprising an open reading frame for (a) ETV2 or ERG, and GATA1;
- (b) ETV2 or ERG, and GATA2;
- (c) ETV2 or ERG, and GFI1;
- or (c) TAL1 and GATA2;
- or
- (ii) one or more recombinant expression viruses suitable for expression, in human pluripotent stem cells, of (a) ETV2 or ERG, and GATA1;
- (b) ETV2 or ERG, and GATA2;
- (c) ETV2 or ERG, and GFI1;
- or (c) TAL1 and GATA2.
Dependencies are inferred from the claim text.