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Updated: Aug 15, 2025
A Culture Method to Maintain Quiescent Human Hematopoietic Stem Cells
Published on: May 17, 2021
Protocol to identify and analyze mouse and human quiescent hematopoietic stem cells using flow cytometry combined
Jiajing Qiu1, Vijay Menon1, Nikolaos Tzavaras2
1Department of Cell, Developmental & Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Abstract:
Mitochondrial membrane potential (MMP) segregates functionally distinct subsets within highly purified hematopoietic stem cells (HSCs). Here, we detail a protocol for FACS isolation of MMP sub-fractions of phenotypically defined mouse and human HSCs. These steps are followed by high-/super-resolution immunofluorescence microscopy of HSCs' lysosomes. While the protocol describes the isolation of quiescent HSCs, which are the most potent subsets, it could also be applied to other HSC subsets. This protocol overcomes some experimental challenges associated with low HSC numbers. For complete details on the use and execution of this protocol, please refer to Liang et al. (2020) and Qiu et al. (2021).
