摘要
应用差酶消化法和反复贴壁法在体外建立奶牛乳腺上皮细胞培养方法,以细胞流式术、免疫组化、免疫印迹、超微结构观察等方法对乳腺上皮细胞特性进行检测,并研究高温热刺激对乳腺细胞超微结构的影响。实验结果表明,运用本方法建立的奶牛乳腺细胞系上皮特性及遗传特征完备;41℃1h的高温热刺激可使乳腺细胞染色质浓缩、线粒体肿胀、空泡化,形成凋亡小体,说明高温可以诱发乳腺细胞凋亡。
A simple method of trypsin /collagenase I alternative digestion and iterative culture flask adherence to discard fibroblasts for bovine mammary cell culture was established in this study. By immunohistochemistry, flow cytometry, western blot, Electron microscopy analysis, the characteristics of bovine mammary cells were investigated in vitro. Effect of hyperthermia on the cell uhrastrnctures was also observed. The results showed that the mammary cells were diploid epithelia with intact 30 pairs chromatins, which could secrete alpha-casein into the medium . After exposed to hyperthermia, the cell condensed chromatin like crescent on the nuclei verges, mitochondria occurred expansion and vacuolization, and apoptotic bodies appeared, which suggested that heat stress could induce apoptosis of the mammary epithelia.
出处
《生物工程学报》
CAS
CSCD
北大核心
2007年第3期471-476,共6页
Chinese Journal of Biotechnology
基金
国家科技部十五重大奶业专项(No.2006BAD04A12)
2005年度江苏省"研究生"创新计划项目资助。~~
关键词
奶牛
乳腺细胞
高温
bovine, mammary cells, heat stress