In:
Stem Cells, Oxford University Press (OUP), Vol. 27, No. 10 ( 2009-10-01), p. 2552-2562
Abstract:
Identification of prevalent specific markers is crucial to stem/progenitor cell purification. Determinants such as the surface antigens CD34 and CD38 are traditionally used to analyze and purify hematopoietic stem/progenitor cells (HSCs/HPCs). However, the variable expression of these membrane antigens poses some limitations to their use in HSC/HPC purification. Techniques based on drug/stain efflux through the ATP-binding cassette (ABC)G2 pump (side population [SP] phenotype) or on detection of aldehyde dehydrogenase (ALDH) activity have been independently developed and distinguish the SP and ALDHBright (ALDHBr) cell subsets for their phenotype and proliferative capability. In this study, we developed a multiparametric flow cytometric method associating both SP and ALDH activities on human lineage negative (Lin−) bone marrow cells and sorted different cell fractions according to their SP/ALDH activity level. We find that Lin−CD34+CD38Low/− cells are found throughout the spectrum of ALDH expression and are enriched especially in ALDHBr cells when associated with SP functionality (SP/ALDHBr fraction). Furthermore, the SP marker identified G0 cells in all ALDH fractions, allowing us to sort quiescent cells regardless of ALDH activity. Moreover, we show that, within the Lin−CD34+CD38−ALDHBr population, the SP marker identifies cells with higher primitive characteristics, in terms of stemness-related gene expression and in vitro and in vivo proliferative potential, than the Lin−CD34+ CD38−ALDHBr main population cells. In conclusion, our study s hows that the coexpression of SP and ALDH markers refines the Lin−CD34+CD38− hematopoietic compartment and identifies an SP/ALDHBr cell subset enriched in quiescent primitive HSCs/HPCs.
Type of Medium:
Online Resource
ISSN:
1066-5099
,
1549-4918
Language:
English
Publisher:
Oxford University Press (OUP)
Publication Date:
2009
detail.hit.zdb_id:
2030643-X
detail.hit.zdb_id:
1143556-2
detail.hit.zdb_id:
605570-9
SSG:
12
