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Logo of arthrestherBioMed Centralbiomed central web sitesearchsubmit a manuscriptregisterthis articleArthritis Research & Therapy
Arthritis Res Ther. 2004; 6(5): R422–R432.
Published online Jul 19, 2004. doi:  10.1186/ar1210
PMCID: PMC546281
Identification of subpopulations with characteristics of mesenchymal progenitor cells from human osteoarthritic cartilage using triple staining for cell surface markers
Stefan Fickert,1,2 Jörg Fiedler,1,3 and Rolf E Brennercorresponding author1,3
1Department of Orthopaedics, University of Ulm, Ulm, Germany
2Department of Orthopaedics, University of Dresden, Dresden, Germany
3Division for Biochemistry of Joint and Connective Tissue Diseases, University of Ulm, Ulm, Germany
corresponding authorCorresponding author.
Jörg Fiedler: joerg.fiedler/at/; Rolf E Brenner: rolf.brenner/at/
Received April 16, 2004; Revisions requested May 19, 2004; Revised May 28, 2004; Accepted June 14, 2004.
We first identified and isolated cellular subpopulations with characteristics of mesenchymal progenitor cells (MPCs) in osteoarthritic cartilage using fluorescence-activated cell sorting (FACS). Cells from osteoarthritic cartilage were enzymatically isolated and analyzed directly or after culture expansion over several passages by FACS using various combinations of surface markers that have been identified on human MPCs (CD9, CD44, CD54, CD90, CD166). Culture expanded cells combined and the subpopulation derived from initially sorted CD9+, CD90+, CD166+ cells were tested for their osteogenic, adipogenic and chondrogenic potential using established differentiation protocols. The differentiation was analyzed by immunohistochemistry and by RT-PCR for the expression of lineage related marker genes. Using FACS analysis we found that various triple combinations of CD9, CD44, CD54, CD90 and CD166 positive cells within osteoarthritic cartilage account for 2–12% of the total population. After adhesion and cultivation their relative amount was markedly higher, with levels between 24% and 48%. Culture expanded cells combined and the initially sorted CD9/CD90/CD166 triple positive subpopulation had multipotency for chondrogenic, osteogenic and adipogenic differentiation. In conclusion, human osteoarthritic cartilage contains cells with characteristics of MPCs. Their relative enrichment during in vitro cultivation and the ability of cell sorting to obtain more homogeneous populations offer interesting perspectives for future studies on the activation of regenerative processes within osteoarthritic joints.
Keywords: cartilage, mesenchymal progenitor cell, osteoarthritis
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