, 58:163 | Cite as

Isolation of osteoprogenitors from murine bone marrow by selection of CD11b negative cells

  • A. Dumas
  • M. A. Le Drévo
  • M. F. Moreau
  • C. Guillet
  • M. F. Baslé
  • D. Chappard
Original Research


Selection of cells having the most osteogenic potential is a strategy used in bone tissue engineering. Preclinical studies using murine bone marrow cells must consider the large amount of hematopoietic cells in the adherent fraction. The aim of this study was to enrich a murine bone marrow cell population with osteoprogenitors by using a simple and reliable method. Bone marrow from C57Bl/6 mice was extracted and cells which adhered onto plastic were expanded in primary culture for 14 days. Immunolabeling of the CD11b surface antigen was performed and the CD11b cell fraction was isolated by FACS. Sorted and unsorted populations were analyzed for gene expression of osteoblast differentiation, alkaline phosphatase (AlkP) activity and matrix mineralization capacities. Selection of CD11b cells increased the number of AlkP+ cells from the plastic adherent fraction from 6.3% ± 0.8 to 56% ± 3.3 with a sevenfold increase in AlkP activity. mRNA analysis revealed a significant increase in the CD11b fraction for Osterix (41-fold), RANKL (17-fold), M-CSF (8-fold) and Runx-2 (8-fold). An osteogenic population was obtained with improved capacities to produce a mineralized extracellular matrix in vitro, independently of the presence of glucocorticoids in the culture medium.


Osteoprogenitor Immunodepletion CD11b Alkaline phosphatase mRNA profiles 



Alkaline phosphatase


Mesenchymal stem cells


Fluorescent activating cell sorting


Cell adhesion molecule



This work was made possible by grants from Contrat de Plan Etat—Région “Pays de la Loire”, INSERM and the Bioregos Program. We thank Laurence Preisser and Marie-Hélène Guilleux from SCCAN Service Commun de Cytométrie et d’Analyses Nucléotidiques, IFR132 for their assistance with RT-PCR quantification.


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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • A. Dumas
    • 1
  • M. A. Le Drévo
    • 1
  • M. F. Moreau
    • 1
  • C. Guillet
    • 2
  • M. F. Baslé
    • 1
  • D. Chappard
    • 1
  1. 1.INSERM, U922 “Remodelage osseux et biomatériaux”, LHEA—Faculté de MédecineAngers CedexFrance
  2. 2.Service Commun de cytométrie et d’analyse nucléotidique (SCCAN)Angers Cedex 9France

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