Cell and Tissue Research

, Volume 313, Issue 3, pp 281–290 | Cite as

STRO-1, HOP-26 (CD63), CD49a and SB-10 (CD166) as markers of primitive human marrow stromal cells and their more differentiated progeny: a comparative investigation in vitro

  • Karina Stewart
  • Peter Monk
  • Susan Walsh
  • Carolyn M. Jefferiss
  • Julie Letchford
  • Jon N. Beresford
Regular Article

Abstract

There is widespread interest in the use of bone marrow stromal cells (BMSC) for tissue reconstruction and repair and for gene therapy. BMSC represent the differentiated progeny of CFU-F, which however comprise a developmentally heterogeneous population as is reflected in the cellular heterogeneity of the cell populations to which they give rise. We have compared the efficacy of monoclonal antibodies recognising a series of stromal antigens, viz. STRO-1, HOP-26, CD49a and SB-10/CD166, as tools for the enrichment of CFU-F prior to culture and as developmental markers for culture-expanded BMSC. In freshly isolated bone marrow mononuclear cells (BMMNC), the proportion of antigen-positive cells was 27%, 46%, 5% and 19% for STRO-1, HOP-26, CD49a and CD166, respectively. All CD49a+ cells co-expressed STRO-1. The degree of CFU-F enrichment obtained with anti-CD49a (~18-fold) by a one-pass immunoselection strategy was significantly greater than that of all other antibodies tested. BMSC expressed higher levels of all antigens investigated (except for HOP-26) compared with BMMNC. Expression of STRO-1 and CD49a remained restricted to a subset of BMSC, whereas all BMSC were SB-10/CD166 positive. Treatment with dexamethasone (10 nM), which promotes the differentiation and further maturation of cells of the osteogenic lineage in this cell culture system, increased the expression of CD49a and HOP-26. The CD49a+ and HOP-26+ fractions of BMSC were further subdivided by dual-labelling with anti-STRO-1 and B4–78 (an antibody recognising the B/L/K isoform of the enzyme alkaline phosphatase), respectively. By using a variety of criteria, the HOP-26 antigen was identified as CD63, a member of the tetraspanin family of proteins thought to modulate integrin compartmentalisation and signalling.

Keywords

CFU-F Bone marrow stromal cells STRO-1 HOP-26/CD63 CD49a Human 

Notes

Acknowledgements

The authors are grateful to the surgeons and theatre staff at The Royal Devon and Exeter Hospital, Exeter, without whom this work would not have been possible.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Karina Stewart
    • 1
  • Peter Monk
    • 2
  • Susan Walsh
    • 1
  • Carolyn M. Jefferiss
    • 1
  • Julie Letchford
    • 1
  • Jon N. Beresford
    • 1
  1. 1.School of Pharmacy and PharmacologyUniversity of BathBathUK
  2. 2.Department of Molecular Biology and BiotechnologyUniversity of SheffieldSheffieldUK

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