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Nordic Bioscience A/S, Denmark Purpose: Hypertrophic of chondrocytes lead to calcification of cartilage matrix which is a pathological event in the early phase of osteoarthritis (OA). The hypertrophic chondrocytes are locate in the deep zone of the articular cartilage and are believed to play an important part in the interaction between subchondral bone and cartilage. The aim of our work was to develop and http://www.selleckchem.com/products/cb-5083.html validate an ex vivo model for OA research, in which hypertrophy chondrocytes could be induced by adding appropriate factors. This model was supposed to be used for examination of cartilage changes related to induction of hypertrophy; aiding the understanding of the role of hypertrophy chondrocytes in OA pathogenesis. Methods: Full depth cartilage explants (from superficial to calcified cartilage) were isolated from bovine femoral condyle and cultured for 21 days with different stimulators: 1) TNF-�� [20ng/ml] + Oncostatin M [10ng/ml], 2) IGF-1 [100ng/ml], 3) bFGF [50ng/ml], and 4) BMP-2[100ng/ml]. Supernatants were collected every two or three days. Safranin O/Fast Green (SaFO) and Tuludine Blue (TuB) staining were used to distinguish the morphology of the explants. Collagen type II turnover was measure by PIINP (formation) and CIIMB (degradation) assay. As a marker of hypertrophy chondrocytes, type X collagen expression was evaluated using reverse transcription followed by real-time polymerase chain reaction (PCR) at day 14 and day 21. Results: Alarmarblue data showed that all explants http://www.selleck.cn/products/PD-0332991.html were alive throughout the 21 day. Both IGF-1 and bFGF groups had enlarged cells at day 7 and 14 (TuB and SaFO stainings). CIIMB and PIINP assay results suggested that both IGF-1 and bFGF had significant (p http://www.selleckchem.com/products/a-1155463.html formation at day 7, but not day 14 and 21. Gene expression of type X collagen stimulated by bFGF increased significantly at day 14 and went down at day 21 compared with control (W/O). The increased type X collagen expression confirmed that bFGF could induce chondrocytes hypertrophic in FDC model. Conclusions: We have developed an explants culture models that include all the different layers of the articular cartilage and were able to induce hypertrophy by stimulating with bFGF. We could detect the gene expression of markers at different culture stage in the ex vivo culture model. We speculate that this model could be used as an important ex vivo model for cartilage degenerative diseases and thereby for investigating the effect of different drug candidates. Disclosures: Pingping Chen, None. MO0067 Bmp2 Is Required for Chondrocyte Maturation and Endochondral Bone Development.Ming Zhang*1, Bing Shu2, Rong Xie2, Meina Wang2, Wei Hou3, Dezhi Tang2, Stephen Harris4, Matthew Hilton5, Yongjun Wang3, Di Chen1.