Dual SP/ALDH functionalities refine the human hematopoietic Lin-CD34+CD38- stem/progenitor cell compartment

Stem Cells. 2009 Oct;27(10):2552-62. doi: 10.1002/stem.186.

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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-ribosyl Cyclase 1 / analysis
  • ADP-ribosyl Cyclase 1 / metabolism
  • 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*
  • Humans
  • Mice
  • Nucleotidyltransferases / analysis
  • Nucleotidyltransferases / metabolism

Substances

  • Antigens, CD34
  • Antigens, Surface
  • Biomarkers
  • Aldehyde Dehydrogenase
  • Nucleotidyltransferases
  • lincosaminide O-nucleotidyltransferase
  • ADP-ribosyl Cyclase 1