The use of multiparameter flow cytometry and cell sorting to characterize native human bone marrow mesenchymal stem cells (MSC).
This chapter describes a method for identification, phenotypic analysis, and cell sorting of rare mesenchymal stem cells (MSCs) from human bone marrow (BM) aspirates. The native BM MSC population is identified based on the CD45(-/low)CD271(+) phenotype. The method consists of three related procedures: Procedure 1 involves a microbead-based pre-enrichment step. Two other procedures describe direct flow cytometric analysis of MSCs following the isolation of the mononuclear cell (MNC) fraction (Procedure 2) or more rapidly, following a simple ammonium chloride-based red cell lysis (Procedure 3). Recently described multi-lineage transcript expression in the CD45(-/low)CD271(+) cells suggests that the native BM MSC fraction could be further subdivided into functionally distinct subpopulations. The present protocols are hoped to help MSC biologists to enter this exciting field of research and to take it forward towards a better understanding of MSC biology in vivo.
Leeds Institute for Rheumatic and Musculoskeletal Medicine, Leeds University, Leeds, UK.
Sally
S
Boxall
Elena
E
Jones
D000068816:Adapalene; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D009281:Naphthalenes / Q000032:analysis; D010641:Phenotype
D009281:Naphthalenes; D000068816:Adapalene; D017493:Leukocyte Common Antigens