人髁突软骨细胞培养方法探讨

Exploration of culture method of human condylar cartilage cell in vitro

目的探讨人胚髁突软骨细胞培养方法。方法分离出人胚髁突软骨组织,分别采用组织块培养法、传统两次酶消化法、改良酶消化法进行髁突软骨细胞原代培养,倒置相差显微镜下进行细胞形态学观察,甲苯胺蓝和HE染色鉴定软骨细胞。结果倒置相差显微镜观察发现,采用组织块培养法,未见细胞从组织块爬出;而传统的两次酶消化法可获细胞数为105/mL,但使用的胶原酶浓度高,消化时间较长;改良的Ⅱ型胶原酶消化联合胰蛋白酶法,对原代髁突软骨细胞分离取得了优良而稳定的效果,可收获细胞数为107/mL。结论改良的Ⅱ型胶原酶消化联合胰蛋白酶法是一种良好的人髁突软骨细胞培养的方法。

Objective To explore the culture method of huaman condylar cartilage cells in vitro. Methods The condylar cartilage tissues were dissected for culture by methods of explant, digestion and modified digestion respectively.The morphology of the cultured condylar cartilage cells were discovered under inversion phase difference microscope, the condylar cartilage cells were identified by HE staining,toluidine blue. Results No cells extended from the explant by way of explant,modified collagenase- II and trypsin digestion method can received stable results in isolating and culturing primary articular chondrocytes, which harvest 10^7 cells. There has a significant differences compared with trypsin digestion. Conclusions Modified digestion culture was a good culturing method for human condylar cartilage cells.