Dual SP/ALDH Functionalities Refine the Human Hematopoietic Lin−CD34+CD38− Stem/Progenitor Cell Compartment

• Published in: Stem cells (Dayton, Ohio) Vol. 27; no. 10; pp. 2552 - 2562
• Main Authors: Pierre‐Louis, Olivier, Clay, Denis, Brunet de la Grange, Philippe, Blazsek, Istvan, Desterke, Christophe, Guerton, Bernadette, Blondeau, Camille, Malfuson, Jean‐Valère, Prat, Marie, Bennaceur‐Griscelli, Annelise, Lataillade, Jean‐Jacques, Le Bousse‐Kerdilès, Marie‐Caroline
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.10.2009
• Subjects: ADP-ribosyl Cyclase 1 - analysis; ADP-ribosyl Cyclase 1 - metabolism; Aldehyde dehydrogenase; Aldehyde Dehydrogenase - analysis; Aldehyde Dehydrogenase - metabolism; Animals; Antigens, CD34 - analysis; Antigens, CD34 - metabolism; Antigens, Surface - analysis; Antigens, Surface - metabolism; Biomarkers - analysis; Biomarkers - metabolism; Cell Cycle - physiology; Cell Dedifferentiation - genetics; Cell Lineage; Cell Separation - methods; Cells, Cultured; Flow Cytometry - methods; Gene Expression Regulation - physiology; Hematopoietic Stem Cell Transplantation - methods; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Hematopoietic stem/progenitor cells; Humans; Mice; Nucleotidyltransferases - analysis; Nucleotidyltransferases - metabolism; Quiescence/G0; Side population
• ISSN: 1066-5099; 1549-4918
• DOI: 10.1002/stem.186

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 shows 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. STEM CELLS 2009;27:2552–2562