作　　者： (黄玉香）; (张建军）; (郭志英）; (谭何新）; (刁勇）;

机构地区： 泉州医学高等专科学校药学院,福建泉州362000 华侨大学生物医学学院,福建泉州362000 中国人民解放军第二军医大学药学院,上海200433

出　　处： 《山东医学高等专科学校学报》 2017年第4期277-280,F0002,F0003,共6页

摘　　要： 目的对大鼠膝关节软骨细胞进行分离和培养,观察大鼠软骨细胞的形态学变化。方法通过酶消化法获得大鼠膝关节软骨细胞并进行传代培养;利用倒置相差显微镜观察各代软骨细胞形态变化;采用CCK-8法检测软骨细胞增殖能力,利用紫外分光光度法获得OD值评价软骨细胞生长趋势;采用细胞免疫荧光法检测Ⅱ型胶原表达情况,在荧光倒置相差显微镜下对软骨细胞进行鉴定。结果在酶消化下可获得高纯度软骨细胞,原代细胞多数呈多角形,细胞形态随传代次数增加有变梭形趋势,细胞活力在第三代之后明显减弱,细胞免疫荧光检测显示胶原Ⅱ高表达。结论成功获得大鼠膝关节软骨细胞并进行形态学的观察比较,P3代前的软骨细胞相对稳定,进行软骨细胞相关研究采用P0至P3代软骨细胞为宜。 Objective Isolated and cultivated chondrocytes, and observed their morphological changes. Methods Isolated cartilage cells from rat knees by enzyme digestion, and cultivated them from one generation to another. The chondrocytes morphology were observed by inverted phase contrast microscope, and the growth of the cells were tested by CCK-8, the collagen II of chondrocytes was identified by cell immunofluorescence method. Results The chondrocytes can be attained by enzyme digestion, most of primary chondrocytes presented polygon, cell viability weakened significantly after P3, and collagen II showed green under fluorescent microscope. Conclusion Attained chondrocytes from rat knees, observed and compared their morphological changes, it was found that they kept relatively stable shapes before P3, it is more suitable to choose the P0 to P3 generations cells for chondrocyte related studies.

关 键 词： 大鼠膝关节 软骨细胞 细胞分离 细胞培养