摘要
采用胶原酶消化法和胰蛋白酶选择性消化法分离、培养和纯化牛乳腺上皮细胞。形态学观察表明,培养的细胞具有典型的上皮细胞形态特征;染色体分析结果表明,培养的细胞具有正常的染色体数目。通过荧光免疫细胞染色方法鉴定了培养的细胞表达上皮细胞特异的角蛋白5和8。该细胞在添加胰岛素、氢化可的松以及羊催乳素的无血清培养液中诱导培养时,用RT-PCR方法检测到了β-酪蛋白基因的转录。这些结果表明,分离培养的细胞是乳腺上皮细胞,这些细胞在诱导培养的条件下能够转录表达β-酪蛋白。
Bovine mammary epithelial cells were isolated and purified by using collagenase digestion of the mammary gland tissue for the primary cell culture, and subsequently using trypsin-selected digestion of the cultured cells for cell purification. Morphological observation revealed that the cultured cells possessed the typical character of epithelial cells. Karyotyping analysis showed the normal chromosome number in cultured cells. The tissue-specific expression of cytokeratin 5 and 8 genes in mammary epithelial cells was identified by immunofluorescent cytochemical staining. The transcription of the beta-casein gene was detected by RT-PCR, when the purified cells were induced with insulin, hydrocortisone and prolactin in the culture medium without serum. The above results indicate that the purified cells are mammary epithelial cells, which can transcript beta-casein mRNA in the induction condition.
基金
内蒙古大学特聘教授科研启动费资助项目(203059)
内蒙古自治区自然科学基金(200408020402)
国家高科技项目(2005AA206110)子课题资助项目
关键词
牛乳腺上皮细胞
细胞培养
核型分析
β-酪蛋白转录
Bovine mammary epithelial cell
Cell culture
Karyotype analysis
Beta-casein transcription
