摘要
背景:课题组在前期研究的基础上,体外分离、培养及鉴定新西兰大白兔距骨软骨细胞。目的:探讨新西兰大白兔距骨软骨细胞体外分离、培养及鉴定的实验方法。方法:课题组采用单纯Ⅱ型胶原酶消化法体外分离新西兰大白兔距骨软骨细胞,进行体外培养,通过形态学观察、甲苯胺蓝染色和Ⅱ型胶原蛋白免疫细胞化学染色对软骨细胞进行鉴定。结果与结论:(1)倒置相差显微镜观察3代以内新西兰大白兔距骨软骨细胞呈多角形或三角形,核为圆形或椭圆形;(2)甲苯胺蓝染色可见新西兰大白兔距骨软骨细胞呈紫蓝色,细胞基质呈蓝色;(3)Ⅱ型胶原蛋白免疫细胞化学染色可见新西兰大白兔距骨软骨细胞胞浆及胞膜出现棕黄色颗粒;(4)实验成功建立了新西兰大白兔距骨软骨细胞体外分离、培养及鉴定体系;实验结果提示3代以内软骨细胞生长良好,保持软骨细胞稳定的生物学特征。
BACKGROUND:Based on the previous studies,the New Zealand rabbit talar chondrocyts were isolated,cultured and identified in vitro.OBJECTIVE:To explore the isolation,culture and identification of New Zealand rabbit talar chondrocyts in vitro.METHODS:The chondrocyts were isolated from the talar cartilage of New Zealand white rabbits by type II collagen enzyme digestion,and then cultured in vitro.The cells were identified by inverted phase contrast microscope,toluidine blue staining and collagen type II immunohistochemical staining.RESULTS AND CONCLUSION:Under the inverted phase contrast microscope,most of passaged chondrocytes presented with polygonal or triangle shape and had round or oval nuclei.Toluidine blue staining showed the hyacinthine chondrocytes and blue cellular matrix.Collagen type II immunohistochemical staining showed that the chondrocytes appeared with brown granules in the cytoplasm and membrane.To conclude,a system that can isolate,culture and identify talar chondrocytes from New Zealand rabbits is successfully established.Talar chondrocytes at passages1-3grow well and have stable biological properties.
作者
王勇平
朱兆金
徐向阳
Wang Yong-ping;Zhu Zhao-jin;Xu Xiang-yang(Department of Orthopedics, Ruijin Hospital of Shanghai Jiao Tong University, Shanghai 200025, China;Department of Orthopedics, First Hospital of Lanzhou University Lanzhou 730000, Gansu Province, China)
出处
《中国组织工程研究》
CAS
北大核心
2017年第28期4475-4480,共6页
Chinese Journal of Tissue Engineering Research
基金
兰州大学第一医院科研基金项目(ldyyyn2017-21)~~
