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US 2025/0333701 A1Published application
Compositions and methods for generating hematopoietic stem cells (hscs)
AssigneesWashington University, Ospedale San Raffaele S.R.L., Fondazione Telethon
InventorsChristopher Sturgeon, Andrea Ditadi
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Abstract
The present disclosure provides methods for generating hematopoietic progenitor cells. In some embodiments, the methods involve an in vitro or ex vivo cell culture model utilizing rentionic acid signaling for producing hematopoietic progenitor cells from pluripotent stem cells.
Claims24
A method of generating a population of hematopoietic progenitor cells from a bulk differentiation culture, the method comprising:
- (i) culturing a population of induced pluripotent stem cells (iPSCs) in a mesoderm differentiation medium comprising a base media supplemented with:
- (a) L-glutamine;
- (b) ascorbic acid;
- (c) monothioglycerol;
- (d) transferrin;
- and
- (e) (1) a bone morphogenic protein (BMP), or (2) an activin receptor-like kinase inhibitor and a GSKβ inhibitor,
- wherein the mesoderm differentiation medium further comprises supplementation with an activator of WNT signaling or a GSKP inhibitor between day 2 and day 3 of differentiation;
- and
- (ii) culturing a population of cells obtained from step (i) in a hematopoietic specification medium comprising a base media supplemented with:
- (a) a FGF,
- (b) a VEGF, and
- (c) a retinoic acid signaling agent,
- from day 3 of differentiation to produce a population of CD34+CD43negCD73negCD184neg hematopoietic progenitor cells.
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