Dual SP/ALDH functionalities refine the human hematopoietic Lin-CD34+CD38- stem/progenitor cell compartment.
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 ALDH(Bright) (ALDH(Br)) 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(+)CD38(Low/-) cells are found throughout the spectrum of ALDH expression and are enriched especially in ALDH(Br) cells when associated with SP functionality (SP/ALDH(Br) fraction). Furthermore, the SP marker identified G(0) cells in all ALDH fractions, allowing us to sort quiescent cells regardless of ALDH activity. Moreover, we show that, within the Lin(-)CD34(+)CD38(-)ALDH(Br) 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(-)ALDH(Br) main population cells. In conclusion, our study shows that the coexpression of SP and ALDH markers refines the Lin(-)CD34(+)CD38(-) hematopoietic compartment and identifies an SP/ALDH(Br) cell subset enriched in quiescent primitive HSCs/HPCs.
Pierre-Louis
Olivier
O
Inserm, U972, Villejuif, France.
Clay
Denis
D
Brunet de la Grange
Philippe
P
Blazsek
Istvan
I
Desterke
Christophe
C
Guerton
Bernadette
B
Blondeau
Camille
C
Malfuson
Jean-Valère
JV
Prat
Marie
M
Bennaceur-Griscelli
Annelise
A
Lataillade
Jean-Jacques
JJ
Le Bousse-Kerdilès
Marie-Caroline
MC
D051997:ADP-ribosyl Cyclase 1 / Q000032:analysis / Q000378:metabolism; D000444:Aldehyde Dehydrogenase / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis / Q000378:metabolism; D000954:Antigens, Surface / Q000032:analysis / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D054337:Cell Dedifferentiation / Q000235:genetics; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation / Q000502:physiology; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009713:Nucleotidyltransferases / Q000032:analysis / Q000378:metabolism
D018952:Antigens, CD34; D000954:Antigens, Surface; D015415:Biomarkers; D000444:Aldehyde Dehydrogenase; D009713:Nucleotidyltransferases; C057535:lincosaminide O-nucleotidyltransferase; D051997:ADP-ribosyl Cyclase 1