大鼠椎间盘软骨终板软骨细胞体外自然退变模型的建立

目的：建立大鼠椎间盘软骨终板软骨细胞体外自然退变模型，为椎间盘退变的机制研究提供载体。方法：酶消化及自然传代法分离培养大鼠椎间盘软骨终板软骨细胞，观察其形态学变化。采用MTT法测定第Ⅲ代软骨细胞的生长曲线。采用免疫细胞化学法检测其特征性Ⅱ型胶原的表达情况，对该模型进行鉴定。结果：大鼠椎间盘软骨终板软骨细胞表达特征性Ⅱ型胶原。第Ⅲ代细胞培养在生长第13天后，细胞分裂能力明显下降，核仁不清，细胞变形明显，以梭形为主，折、旋光性弱，细胞间隙变大，轮廓增强，胞浆内可见空泡、脂滴；Ⅱ型胶原合成明显下降，细胞增殖率显著降低；呈现自然退变过程。结论：建立椎间盘软骨终板软骨细胞体外自然退变模型，为椎间盘退变机制研究提供较好的细胞学基础。

An in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats

OBJECTIVE: To set up a natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats in order to offer an appropriate carrier for the study on mechanism of intervertebral disc degeneration. METHODS: The method of enzyme digestion combined with natural subculture was used to set up the in vitro natural degeneration model of chondrocytes derived from the endplate of intervertebral disc of rats. The morphological appearances and microstructures of the chondrocytes of different generations were observed. The expression of collagen II in chondrocytes was detected by immunocytochemical method. RESULTS: The chondrocytes derived from the endplate of intervertebral disc expressed collagen II. After 13 days of culture, the chondrocytes of generation III showed that the ability of cell division descended, the nucleoli became unclear, the cells deformed obviously, fusiform shape with weak optical activity appeared, and the intercellular space was enlarged. There were vacuoles and lipid droplets in cytoplasm. The synthesis of collagen II, as well as the cell proliferation rate, descended notably. All results showed the natural degeneration process of the chondrocytes. CONCLUSION: The in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats was successfully established. This can offer the cytological basis for study on the mechanism of intervertebral disc degeneration.