奶山羊乳腺上皮细胞的分离、培养及鉴定

Characterization and culture of isolated primary dairy goat mammary gland epithelial cells

摘要:

应用组织块培养法高密度培养、连续传代法建立西农萨能奶山羊乳腺上皮细胞体外培养体系，通过生长曲线绘制、核型分析、免疫荧光染色 (角蛋白、上皮膜抗原、波形蛋白、β-酪蛋白)、油红染色及β-酪蛋白基因的RT-PCR分析进行培养细胞鉴定。实验结果表明细胞生长曲线为典型的S型，染色体数目众数为60，细胞角蛋白、上皮膜抗原、波形蛋白、β-酪蛋白表达均呈阳性，油红染色后可见细胞质内的脂滴，且细胞表达酪蛋白mRNA。说明运用本方法培养的细胞为正常的乳腺上皮细胞，并具有一定的泌乳功能。

Abstract:

Based on the in vitro culturing system developed for epithelial cells in mammary gland of Xinong Saanen dairy goats using tissue explants 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, β-casein), oil red staining and RT-PCR of β-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 β-casein was detected, the cytoplasmic lipid droplets were observed following the oil red staining, the cultured cells expressed the mRNA of β-casein. In conclusion, the current in vitro culturing system can obtain the normal mammary gland epithelial cells with the function of secretion.