西藏小型猪肋软骨细胞的分离培养及鉴定  

Isolation and identification of Tibetan miniature pig rib chondrocytes

作　　者：刘薇[1] 史俊文 岳敏[1] 张婷婷 唐华[1] 袁进[1] 肖东[1] 顾为望[1] 

机构地区：[1]南方医科大学比较医学研究所暨实验动物中心 [2]肿瘤研究所,广州510515

出　　处：《中国比较医学杂志》2013年第3期1-3,F0002,I0001,共5页Chinese Journal of Comparative Medicine

基　　金：国家863项目(No.2011AA100304);广州市科技计划项目(No.2010U1-E00811);国家自然科学基金项目(No.81172587);广东省科研条件建设项目(No.1021010200004)

摘　　要：目的分离培养西藏小型猪肋软骨细胞,并对其进行鉴定。方法实验采用3日龄西藏小型猪,二步消化法分离培养西藏小型猪肋软骨细胞(PCCs),同样的方法分离培养小鼠的肋软骨细胞(MCCs),观察两个物种软骨细胞的形态,并分别以西藏小型猪胚胎成纤维细胞(PEFs)和小鼠胚胎成纤维细胞(MEFs)为阴性对照,对所分离培养的软骨细胞进行甲苯胺蓝染色鉴定以及胶原蛋白Ⅱ、蛋白聚糖的免疫荧光鉴定。结果成功分离培养西藏小型猪和小鼠肋软骨细胞,两种细胞的形态有差别,所分离培养的两个物种的软骨细胞均呈甲苯胺蓝染色阳性,胶原蛋白Ⅱ、蛋白聚糖免疫荧光阳性。结论成功建立了西藏小型猪肋软骨细胞分离培养及鉴定的方法。Objective Isolation and culture of Tibetan miniature pig rib chondrocytes （PCCs）, followed by identifying it. Methods A 3-day-old Tibetan miniature pig was used in this experiment. Both PCCs and mouse rib ehondrocytes （MCCs） were isolated by two-step digestion method. The isolated chondrocytes were identified by toluidine blue staining and immunofluoreseence for collagen Ⅱ and aggrecan. Tibetan miniature pig embryonic fihroblasts （PEFs） and mouse embryonic fibroblasts （MEFs） were employed as a negative control. Results PCCs and MCCs were successfully isolated and cultured, and demonstrated the different cell morphology, whereas toluidine blue staining and immunofluorescence staining confirmed the chondrocytes characteristics of isolated PCCs. Conclusion The method for isolating, culturing and identifying PCCs of Tibetan miniature pig was successfully established.

关 键 词：西藏小型猪 软骨细胞 甲苯胺蓝 胶原蛋白Ⅱ 蛋白聚糖 

分 类 号：R-332[医药卫生]