体外兔骨髓基质干细胞软骨方向诱导

Introduced transformation of bone mesenchymal stem cells of rabbits to chondrocytic differentiation in vitro

目的 探讨兔骨髓基质干细胞 (bonemesenchymalstemcell ,BMSC)体外定向软骨样分化的有效诱导条件及相关生物学特性。方法 选用 8周新西兰大白兔 ,于胫骨或股骨抽取骨髓 ,经梯度离心、培养、扩增 ,取第 3代BMSC按一定比例种植于 6孔板中 ,并加入含有转化生长因子 β1、地塞米松和维生素C的培养液。于不同时间点取材固定 ,甲苯胺蓝染色和II型胶原免疫组化鉴别。结果 在诱导液作用下 ,BMSC增殖速度逐渐减慢 ,其形态由梭形变为圆形 ,但合成类软骨基质增多 ,细胞外基质呈甲苯胺蓝异染性和II型胶原免疫组化阳性。结论 在本实验条件下 ,获取的兔BMSC生长稳定 ,增殖快 。

Objective To study the induced conditions and relativ e biological properties of bone mesenchymal stem cell (BMSC) of rabbit into chondr ogenesis in vitro. Methods Bone marrow was harvested from t ibia s or femurs of 8-week-old New Zealand white rabbits. After gradient centrifug ation,cultivation,amplification, the 3rd- passaged BMSC were implanted in six- hole-plate according to a certain proportion and induced by chondrogenic induce rs including transforming growth factor-β 1, dexamethasone and vitamin C.Chon drocytes were selected and fixed at different time. The features of chondrocytes were identified by toluidine blue staining and collagen types II immunohistoche mical assay. Results During the introduction, the growth rate o f BMSC became slowly and the shape changed from fusiform to round,but the synthe sis of cartilage matrix increased,and positive results were showed on the to luidine blue staining and collagen types II immunohistochemical staining. Conclusion Rabbit BMSC ,obtained under the experimental conditions,d evelopes stably, proliferates rapidly and is differentiated successfully into ch ondrocytes by induction in vitro.