Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro

✍ Scribed by M. Jakob; O. Démarteau; D. Schäfer; B. Hintermann; W. Dick; M. Heberer; I. Martin

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 570 KB
• Volume: 81
• Category: Article
• ISSN: 0730-2312
• DOI: 10.1002/1097-4644(20010501)81:2<368::aid-jcb1051>3.0.co;2-j

✦ Synopsis

Adult human articular chondrocytes were expanded in a medium with 10% serum (CTR) or further supplemented with different mitogens (i.e., EGF, PDGFbb, FGF-2, TGFb1, or FGF-2/TGFb1). Cells were then induced to redifferentiate in 3D pellets using serum-supplemented medium (SSM), serum-free medium (SFM), or SFM supplemented with factors inducing differentiation of chondroprogenitor cells (i.e., TGFb1 and/or dexamethasone). All factors tested during expansion enhanced chondrocyte proliferation and dedifferentiation, as assessed by the mRNA ratios of collagen type II to type I (CII/CI) and aggrecan to versican (Agg/Ver), using real-time PCR. FGF-2/TGFb1expanded chondrocytes displayed the lowest doubling times, CII/CI and Agg/Ver ratios, averaging, respectively, 50, 0.2 and 15% of CTR-expanded cells. Redifferentiation in pellets was more ef®cient in SFM than SSM only for EGF-, PDGFbb-or FGF-2-expanded chondrocytes. Upon supplementation of SFM with TGFb and dexamethasone (SFM TD), CII/CI ratios decreased 4.4-fold for EGF-and PDGFbb-expanded chondrocytes, but increased 96-fold for FGF-2/TGFb1expanded cells. Chondrocytes expanded with FGF-2/TGFb1 and redifferentiated in SFM TD expressed the largest mRNA amounts of CII and aggrecan and generated cartilaginous tissues with the highest accumulation of glycosaminoglycans and collagen type II. Our results provide evidence that growth factors during chondrocyte expansion not only in¯uence cell proliferation and differentiation, but also the cell potential to redifferentiate and respond to regulatory molecules upon transfer into a 3D environment.