奶牛原代乳腺上皮细胞的培养及β酪蛋白mRNA的表达

Culture of Dairy Cow Primary Mammary Epithelial Cells and Expression of β-casein mRNA

本试验旨在建立原代乳腺上皮细胞系的体外培养方法,并进行β酪蛋白mRNA的表达验证。取新鲜泌乳期的乳腺组织,采用组织块法培养纯化原代乳腺上皮细胞,利用显微镜观察细胞形态并进行细胞生长计数,采用实时荧光定量PCR技术检测β酪蛋白mRNA表达。结果显示,纯化培养的原代乳腺上皮细胞集聚成岛屿状生长,具有典型的铺路石和鹅卵石形状,细胞生长曲线呈"S"形,符合一般细胞的生长规律,并成功表达β酪蛋白mRNA。综上所述,本研究采用组织块法成功培养出具有正常生理功能的奶牛原代乳腺上皮细胞,为后续的乳腺上皮细胞功能研究提供了良好的细胞试验模型。

The aim of this study was to construct the culture method of primary mammary epithelial cells in vitro using mammary tissue and to test expression ofβ-casein mRNA in mammary gland epithelial cells.The fresh lactating mammary gland tissue was obtained,microscope observation was used for cell morphology and cell growth counting,and the Real-time PCR method was used for detection ofβ-casein mRNA expression.The results showed that purified primary mammary gland epithelial cells formed island-like and grew with typical shape of slabstone and cobblestone;Primary mammary gland epithelial cells growth curve was＂S＂type and accorded with rule of general cell growth.Mammary epithelial cells expressedβ-casein mRNA successfully.In conclusion,the primary mammary epithelial cells with normal physiology function had been established successfully.The study provided favourable cell model for further research of the functions of mammary epithelial cells.