title: Effects of transforming growth factor-β subtypes on in vitro cartilage production and mineralization of human bone marrow stromal-derived mesenchymal stem cells.
abstract: Human bone marrow stromal-derived mesenchymal stem cells (hBMSCs) will differentiate into chondrocytes in response to defined chondrogenic medium containing transforming growth factor-β (TGFβ). Results in the literature suggest that the three mammalian subtypes of TGFβ (TGFβ1, TGFβ2 and TGFβ3) provoke certain subtype-specific activities. Therefore, the aim of our study was to investigate whether the TGFβ subtypes affect chondrogenic differentiation of in vitro cultured hBMSCs differently. HBMSC pellets were cultured for 5 weeks in chondrogenic media containing either 2.5, 10 or 25 ng/ml of TGFβ1, TGFβ2 or TGFβ3. All TGFβ subtypes showed a comparable dose-response curve, with significantly less cartilage when 2.5 ng/ml was used and no differences between 10 and 25 ng/ml. Four donors with variable chondrogenic capacity were used to evaluate the effect of 10 ng/ml of either TGFβ subtype on cartilage formation. No significant TGFβ subtype-dependent differences were observed in the total amount of collagen or glycosaminoglycans. Cells from a donor with low chondrogenic capacity performed equally badly with all TGFβ subtypes, while a good donor overall performed well. After addition of β-glycerophosphate during the last 2 weeks of culture, the expression of hypertrophy markers was analysed and mineralization was demonstrated by alkaline phosphatase activity and alizarin red staining. No significant TGFβ subtype-dependent differences were observed in expression collagen type X or VEGF secretion. Nevertheless, pellets cultured with TGFβ1 had significantly less mineralization than pellets cultured with TGFβ3. In conclusion, this study suggests that TGFβ subtypes do affect terminal differentiation of in vitro cultured hBMSCs differently.
authors.affiliation: Department of Otorhinolaryngology and Head and Neck Surgery, Erasmus MC, University Medical Centre Rotterdam, The Netherlands.
authors.forename: F L J
authors.initials: FL
authors.lastname: Cals
authors.forename: C A
authors.initials: CA
authors.lastname: Hellingman
authors.forename: W
authors.initials: W
authors.lastname: Koevoet
authors.forename: R J
authors.initials: RJ
authors.lastname: Baatenburg de Jong
authors.forename: G J V M
authors.initials: GJ
authors.lastname: van Osch
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D002356:Cartilage / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods; D019902:Chondrocytes / Q000166:cytology; D024121:Collagen Type X / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005994:Glycerophosphates / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*; D053781:Transforming Growth Factor beta2 / Q000378:metabolism*; D053782:Transforming Growth Factor beta3 / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D024121:Collagen Type X; D005994:Glycerophosphates; C508960:TGFB1 protein, human; C509041:TGFB2 protein, human; C509044:TGFB3 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D053781:Transforming Growth Factor beta2; D053782:Transforming Growth Factor beta3; D042461:Vascular Endothelial Growth Factor A; C031463:beta-glycerophosphoric acid