兔骨髓基质干细胞体外定向分化为软骨细胞

Inducing Rabbits Bone Mesenchymal Stem Cells into Chondrocytes in Vitro

目的体外诱导兔骨髓基质干细胞(BMSC)向软骨细胞定向分化,并对分化后的细胞进行鉴定.方法选用16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.结果诱导的软骨样细胞甲苯氨蓝异染性;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,并成功地诱导其向软骨细胞表型转化.

Objective To induce bone marrow mesenchymal stem cells （BMSCs） or bone marrow stroma stem cell （MSCs） to differentiate directionally towards chondrocytes in vitro and then identify the differentiated cells. Methods Bone marrow was harvested from the iliac bone of 16-week-old Japanese white rabbits. After gradient centrifugation, cultivation, amplification, the 3rd - passaged BMSC were implanted in six-hole-plate according to a certain proportion and induced by chondrogenic inducers including transforming growth factor-131, dexamethasone and vitamin C. Chondrocytes were selected and fixed at different time. The features of chon-drocytes were identified by toluidine blue staining and collagen types II immunohistochemical assay. Results The structure of cellular cartilage from BMSCs was uniformly positive of toluidine blue staining and collagen types II immunohistochemical staining. Gonclusion Rabbit BMSC, obtained under the experimental conditions, de-velops stably, proliferates rapidly and is differentiated successfully into chondrocytes by induction in vitro.