Protocol to maintain single functional mouse hematopoietic stem cells in vitro without cell division

This protocol details the isolation and in vitro maintenance of single hematopoietic stem cells (HSCs) in the absence of the bone marrow niche. The HSCs do not divide over 7 days and fully retain their long-term functional capacity in transplantation assays. Following hibernation culture, HSCs can be used to study quiescence exit and can be genetically manipulated as single cells prior to division. For complete details on the use and execution of this protocol, please refer to Oedekoven et al. (2021).