title: CD14-negative isolation enhances chondrogenesis in synovial fibroblasts.
abstract: Synovial membrane has been shown to contain mesenchymal stem cells. We hypothesized that an enriched population of synovial fibroblasts would undergo chondrogenic differentiation and secrete cartilage extracellular matrix to a greater extent than would a mixed synovial cell population (MSCP). The optimum doses of transforming growth factor beta 1 (TGF-beta1) and insulin-like growth factor 1 (IGF-1) for chondrogenesis were investigated. CD14-negative isolation was used to obtain a porcine cell population enriched in type-B synovial fibroblasts (SFB) from an MSCP. The positive cell surface markers in SFB were CD90, CD44, and cadherin-11. SFB and MSCP were cultured in the presence of 20 ng/mL TGF-beta1 for 7 days, and SFB were demonstrated to have higher chondrogenic potential. Further dose-response studies were carried out using the SFB cells and several doses of TGF-beta1 (2, 10, 20, and 40 ng/mL) and/or IGF-1 (1, 10, 100, and 500 ng/mL) for 14 days. TGF-beta1 supplementation was essential for chondrogenesis and prevention of cell death, whereas IGF-1 did not have a significant effect on the SFB cell number or glycosaminoglycan production. This study demonstrates that the CD14-negative isolation yields an enhanced cell population SFB that is more potent than MSCP as a cell source for cartilage tissue engineering.
authors.affiliation: Department of Orthopaedics, Alpert Medical School of Brown University and Center for Restorative and Regenerative Medicine, Providence VA Medical Center and Brown University, Providence, Rhode Island 02903, USA.
authors.forename: Bahar
authors.initials: B
authors.lastname: Bilgen
authors.forename: Yuexin
authors.initials: Y
authors.lastname: Ren
authors.forename: Ming
authors.initials: M
authors.lastname: Pei
authors.forename: Roy K
authors.initials: RK
authors.lastname: Aaron
authors.forename: Deborah McK
authors.initials: DM
authors.lastname: Ciombor
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D018960:Hyaluronan Receptors / Q000378:metabolism*; D013552:Swine; D013583:Synovial Membrane / Q000166:cytology* / Q000187:drug effects / Q000503:physiopathology*; D023822:Tissue Engineering / Q000379:methods; D053773:Transforming Growth Factor beta1 / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D018960:Hyaluronan Receptors; D053773:Transforming Growth Factor beta1