Wang Zhen, Luo Jun, Wang Wei, Zhao Wangsheng, Lin Xianzi
College of Animal Science and Technology, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling 712100, China.
Sheng Wu Gong Cheng Xue Bao. 2010 Aug;26(8):1123-7.
Based on the in vitro culturing system developed for epithelial cells in mammary gland of Xinong Saanen dairy goats using tissue explant culture, high density cultivation, and continuous passaging, the cultured epithelial cells were evaluated by growth curve fitting, karyotype analysis, immunofluorescence staining (keratin, epithelial membrane antigen (EMA), vimentin, beta-casein), oil red staining and RT-PCR of beta-casein gene. The results showed that the growth of epithelial cells with the model number of chromosome of 60 demonstrated a typical 'S' shape curve, the positive gene expression of keratin, EMA, vimentin and beta-casein was detected, the cytoplasmic lipid droplets were observed following the oil red staining, the cultured cells expressed the mRNA of beta-casein. In conclusion, the current in vitro culturing system can obtain the normal mammary gland epithelial cells with the function of secretion.
基于利用组织块培养、高密度培养和连续传代技术建立的西农萨能奶山羊乳腺上皮细胞体外培养体系,通过生长曲线拟合、核型分析、免疫荧光染色(角蛋白、上皮膜抗原(EMA)、波形蛋白、β-酪蛋白)、油红染色及β-酪蛋白基因的RT-PCR对培养的上皮细胞进行评估。结果显示,染色体数目为60条的上皮细胞生长呈典型的“S”形曲线,检测到角蛋白、EMA、波形蛋白和β-酪蛋白的基因表达呈阳性,油红染色后观察到细胞质脂滴,培养细胞表达β-酪蛋白的mRNA。综上所述,当前的体外培养体系能够获得具有分泌功能的正常乳腺上皮细胞。