Maintenance of hematopoietic stem cells (HSCs) in vitro has been believed to be difficult due to a lack of complete understanding of HSC quiescence maintained by the niche.Recent evidence suggests that in vitro maintenance of human and mouse long-term HSCs (LT-HSCs)is possible through dual inhibition (2i) of both GSK-3 and mTOR in the absence of cytokines,serum,or feeder cells.