人胚颞颌关节软骨细胞培养及生物学特性研究

The Culture of TMJ Condylar Cartilage Cells and Study on Their Biological Behavoirs in Vitro

采用机械分离及胰蛋白酶和Ⅱ型胶原酶联合消化的方法，简便、快速地获得大量成活率高的人ＴＭＪ软骨细胞。在ＤＭＥＭ培养基中进行原代和传代培养，并对原代培养软骨细胞进行了组化及免疫组化鉴定，相差显微镜、光镜及超微结构的观察。结果显示，光镜观察，原代培养细胞呈多角形，单层排列，电镜可见细胞内有丰富的粗面内质网及线粒体，细胞呈多极性，表面有突起。ＴＭＪ软骨细胞免疫组化Ⅱ型胶原染色阳性。甲苯胺蓝染色细胞质内有大量异染基质颗粒，说明本实验培养的ＴＭＪ软骨细胞保持了体内的基本特征。

To investigate the behavoir of TMJ condylar cartilage cells in vitro, the mandibular condylar cartilage cells were harvested from a 5 month old human fetus by dissection and sequential digestion with 0.25% trypsin and 0.2% collagenase (type Ⅱ). The isolated cells were cultured in DMEM medium and identified by histochemical and immunohistochemical methods. Cell proliferation, morphology and ultrastructure were observed by phase contrast microscope, cytologic staining and electronic microscope. In primarily cultured cells, polygonal chondroblast like cells dominated and they were confirmed by the positive result of immunohistochemical examination for type Ⅱ collagen and Toludin blue staining. In conclusion, the TMJ cells in this culture system kept their phenotype in vivo.