Article
Reduced CD166 expression in mesenchymal stromal cells from osteoarthritic donors
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Authors
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A. Jacobi
- University Hospital Carl Gustav Carus, Department of Orthopedics, Dresden, Germany
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H. Friedrich
- University Hospital Carl Gustav Carus, Department of Orthopedics, Dresden, Germany
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J. Rauh
- University Hospital Carl Gustav Carus, Department of Orthopedics, Dresden, Germany
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C. Vater
- University Hospital Carl Gustav Carus, Department of Orthopedics, Dresden, Germany
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K.-P. Günther
- University Hospital Carl Gustav Carus, Department of Orthopedics, Dresden, Germany
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M. Stiehler
- University Hospital Carl Gustav Carus, Department of Orthopedics, Dresden, Germany
| Published: | October 21, 2010 |
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Outline
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Objective: Osteoarthritis (OA) as a common degenerative joint disease denotes a major socioeconomic problem. Cell-based strategies for the therapy of a variety of musculoskeletal disorders have received increasing attention. In this context, multipotent mesenchymal stromal cells (MSCs) represent an attractive cellular component for tissue engineering strategies (Stiehler et al., Adv Exp Med Biol, 2006.). Analysis of cellular surface markers is essential for the identification and detailed characterization of MSCs (Dominici et al., Cytotherapy, 2006.). To assess whether advanced-stage OA affects MSC phenotype we investigated cellular surface marker expression patterns of MSCs from osteoarthritic in contrast to healthy donors using fluorescence-activated cytometric sorting (FACS) analysis.
Methods: MSCs were isolated from bone marrow aspirates (50 cc each) obtained from the pelvic compartment of n=14 advanced-stage idiopathic hip osteoarthritic (Kellgren and Lawrence grade 3 or 4, mean age 67±6 years) and n=15 age-matched (61±4 years) healthy donors by Ficoll gradient-separation and plastic adherence. OA was objectified radiologically (only OA group) and by use of algofunctional indices (i.e. Western Ontario McMaster Universities OA index, Harris Hip Score, EuroQol-5D). The study was approved by the local institutinal review board (protocol # EK203082008).
Cells were maintained in Dulbecco's Modified Eagle Medium supplemented with 10% fetal calf serum at 5% CO2. To assess expression of CD34, CD45, CD14, CD49, CD73, CD90, CD106, CD146, CD166 (all purchased from BD Biosciences), CD105 (AbD Serotec) and STRO-1 (Biolegend), FACS analysis was performed using passage 2 MSCs on a BDTM LSRII flow cytometer. Mean flourescence intensities and percentages of positive viable cells were determined.
Results were considered significant for two-sided p-values <0.05 based on all pairwise comparisons.
Ergebnisse und Schlussfolgerungen: Expression levels of CD73, CD90, and STRO-1 were elevated in respect of the negative control cellular surface antigens CD14, CD34, and CD45 in MSCs derived from both osteoarthritic and healthy donors (p<0.0001). Interestingly, MSCs from OA patients showed decreased levels of CD166 expression compared to cells from healthy donors (p<0.05).
The surface marker pattern of MSCs isolated from OA patients is generally comparable to that of MSCs from healthy donors. However, the observed reduced CD166 expression levels in OA-MSCs warrants further investigation. These data will help to facilitate the application of autologous cell-based strategies for musculoskeletal tissue regeneration.
