Dual SP/ALDH Functionalities Refine the Human Hematopoietic LinCD34+CD38 Stem/Progenitor Cell Compartment§

Authors

  • Olivier Pierre-Louis,

    1. Inserm, U972, Villejuif, France
    2. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    3. IFR89, Institut André Lwoff, Villejuif, France
    Search for more papers by this author
  • Denis Clay,

    1. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    2. IFR89, Institut André Lwoff, Villejuif, France
    Search for more papers by this author
  • Philippe Brunet de la Grange,

    1. Inserm, U972, Villejuif, France
    2. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    3. IFR89, Institut André Lwoff, Villejuif, France
    Search for more papers by this author
  • Istvan Blazsek,

    1. Inserm, U972, Villejuif, France
    2. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    3. IFR89, Institut André Lwoff, Villejuif, France
    Search for more papers by this author
  • Christophe Desterke,

    1. Inserm, U972, Villejuif, France
    2. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    3. IFR89, Institut André Lwoff, Villejuif, France
    Search for more papers by this author
  • Bernadette Guerton,

    1. Inserm, U972, Villejuif, France
    2. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    3. IFR89, Institut André Lwoff, Villejuif, France
    Search for more papers by this author
  • Camille Blondeau,

    1. Service d'Orthopédie, Centre Hospitalier Robert Ballanger, Aulnay-Sous-Bois, France
    Search for more papers by this author
  • Jean-Valère Malfuson,

    1. Centre de Transfusion Sanguine des Armées, Hôpital Percy, Clamart, France
    Search for more papers by this author
  • Marie Prat,

    1. Centre de Transfusion Sanguine des Armées, Hôpital Percy, Clamart, France
    Search for more papers by this author
  • Annelise Bennaceur-Griscelli,

    1. Inserm, U935, Villejuif, France
    2. Hôpital Paul Brousse, Villejuif, France
    Search for more papers by this author
  • Jean-Jacques Lataillade,

    Corresponding author
    1. Centre de Transfusion Sanguine des Armées, Hôpital Percy, Clamart, France
    • Jean-Jacques Lataillade, Centre de Transfusion Sanguine des Armées, BP 410, Clamart 92140, France

      Marie-Caroline Le Bousse-Kerdilès, Inserm U972, 14 Avenue Paul-Vaillant Couturier, Villejuif Cedex 94807, France

    Search for more papers by this author
    • Telephone: 33-1-41-46-72-60; Fax: 33-1-46-38-82-87

    • Jean-Jacques Lataillade and Marie-Caroline Le Bousse-Kerdilès equally coordinated the work.

  • Marie-Caroline Le Bousse-Kerdilès

    Corresponding author
    1. Inserm, U972, Villejuif, France
    2. Université Paris-Sud, Institut André Lwoff, Villejuif, France
    3. Hôpital Paul Brousse, Villejuif, France
    • Jean-Jacques Lataillade, Centre de Transfusion Sanguine des Armées, BP 410, Clamart 92140, France

      Marie-Caroline Le Bousse-Kerdilès, Inserm U972, 14 Avenue Paul-Vaillant Couturier, Villejuif Cedex 94807, France

    Search for more papers by this author
    • Jean-Jacques Lataillade and Marie-Caroline Le Bousse-Kerdilès equally coordinated the work.

    • Telephone: 33-1-45-59-53-03; Fax: 33-1-47-26-03-19


  • Author contributions: O.P.-L.: study conception, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; D.C.: flow cytometry experiment conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; P.B.G.: NOD-SCID experiment conception and design, manuscript rewriting, final approval of manuscript; I.B.: data analysis and interpretation, manuscript writing; C.D.: assembly of data, statistical analyses; B.G.: bone marrow cell isolation; C.B.: provision of bone marrow sample; J.V.M.: participation in the research design, provision of bone marrow sample, and final approval of manuscript; M.P.: bone marrow sample provision, final approval of manuscript; A.B.-G.: discussion and manuscript revision; J.-J.L.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript; M.-C.L.B.-K.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLS EXPRESS July 30, 2009.

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 LinCD34+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 LinCD34+CD38ALDHBr 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 LinCD34+ CD38ALDHBr main population cells. In conclusion, our study shows that the coexpression of SP and ALDH markers refines the LinCD34+CD38 hematopoietic compartment and identifies an SP/ALDHBr cell subset enriched in quiescent primitive HSCs/HPCs. STEM CELLS 2009;27:2552–2562

Ancillary