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

Miriam Belmonte; David G Kent

doi:10.1016/j.xpro.2021.100927

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

STAR Protoc. 2021 Nov 1;2(4):100927. doi: 10.1016/j.xpro.2021.100927. eCollection 2021 Dec 17.

Authors

Miriam Belmonte^{1

2}, David G Kent^{1

2

3}

Affiliations

¹ Wellcome MRC Cambridge Stem Cell Institute, University of Cambridge, Hills Road, Cambridge CB2 0XY, UK.
² Department of Haematology, University of Cambridge, Cambridge CB2 0XY, UK.
³ York Biomedical Research Institute, Department of Biology, University of York, York YO10 5DD, UK.

Abstract

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).

Keywords: Cell culture; Flow Cytometry/Mass Cytometry; Immunology; Microscopy; Single Cell; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods*
Cell Division
Cells, Cultured
Culture Media
Hematopoietic Stem Cells / cytology*
Mice
Single-Cell Analysis

Substances

Culture Media

Abstract

Publication types

MeSH terms

Substances

Grants and funding