中国组织工程研究与临床康复2011，Vol.15Issue(6)：963-966,4.DOI:10.3969/j.issn.1673-8225.2011.06.004
兔骨髓间充质干细胞培养及向成软骨细胞的诱导分化
Culture and differentiation of rabbit bone marrow mesenchymal stem cells into chondrocytes
摘要
Abstract
BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) do not have ideal specific surface markers, and there is no unified standard to identify BMSCs. Most identification depends on its morphological levels, functional characteristics and differentiational phenotype occurred during the culture.OBJECTIVE: To culture, am plify and identify rabbit BMSCs in vitro, and observe the feasibility of differentiation into chondrocytes.METHODS: Rabbit BMSCs were extracted through the density gradient centrifugation. Cell morphology was observed using the inverted microscope. Cells were cultured in α-MEM medium containing 1O％ fetal bovine serum, and cells were counted daily to observe cell growth speed. Expression of CD45, CD29, CD44 on PO and P3 BMSCs was detected using flow cytometry.Well-grew rabbit BMSCs at P3 were obtained and induced using special media supplemented with 10％ fetal bovine serum,transforming growth factor (TGF)-1 10 μg/L, insulin-like growth factor (IGF)- Ⅰ 110 μg/L, transferring 6.25 mg/L, dexamethasone 10 mmol/L and vitamin C 0.05mmol/L. Morphological changes were observed using the inverted microscope. Specificity of cartilage collagen type Ⅱ was detected using immunohistochemical method.RESU LTS AND CONCLUSION: The cultured cells in the fusiform long, fibroblast growth of samples. The cells grow rapidly. Flow cytometry results showed that for the PO cells about 62.4％ of the cells expressed CD29, about 60.3％ of the cells expressed CD44, 2.7g％ of the cells expressed CD45, and for the P3 cells, about 99.9％ of the cells expressed CD29, about 99.6％ of the cells expressed CD44, 2.62％ of the cells expressed CD45. After the BMSCs in induction, the ability of expansion was decreased obviously, and the morphology changes after 21 days, and the collagen type Ⅱ become dyeing. Results suggest that BMSCs can be successfully isolated and amplified by density gradient centrifugation. The third passage of BMSCs shows high purity. Under appropriate conditions, the BMSCs have the potential of differentiating itself into chondrocytes.关键词
软骨细胞/骨髓间充质干细胞/流式细胞术/表面抗原/诱导分化分类
医药卫生引用本文复制引用
郭璇,霍然,吕仁荣,林莉..兔骨髓间充质干细胞培养及向成软骨细胞的诱导分化[J].中国组织工程研究与临床康复,2011,15(6):963-966,4.基金项目
青年科学研究基金(2004BS02009). （2004BS02009）
