动物营养学报Issue(8)：2544-2550,7.DOI:10.3969/j.issn.1006-267x.2015.08.028
奶牛乳腺上皮细胞系的培养与鉴定
Culture and Identification of the Bovine Mammary Epithelial Cell Line
摘要
Abstract
To establish a functional model of bovine mammary epithelial cell line for the study of lactation reg-ulation, pathogenesis of mastitis, this research applied tissue culture to isolate and culture bovine mammary epi-thelial cells; the limiting dilution method to purify bovine mammary epithelial cells; methyl thiazolyl tetrazoli-um ( MTT) method was used to identify whether the growth curve of bovine mammary epithelial cells was‘S’ type or not;cytokeratin 18 immunofluorescence was observed to prove that the cells were epithelial type;kary-otype analysis was carried out on bovine mammary epithelial cells in passages 10 and 20. The results showed as follows:1) bovine mammary epithelial cells could be successfully cultured and passaged 20 generations by tis-sue culture method. 2) At 5 to 6 days of cultivation, the fibroblast cells proliferated rapidly and a small amount of epithelial cells divided. At 8 days of cultivation, bovine mammary epithelial cells proliferated rapidly and formed island-like colonies, and grew like‘cobblestone’ and‘slabstone’ as monolayer. 3) Bovine mammary epithelial cells showed positive reaction against cytokeratine 18 antigen. 4) At passages 10 and 20 of cultiva-tion, the cell chromosome number was 60 and the cells had normal diploid karyotype. In conclusion, tissue culture cells are able to obtain stable, functional bovine mammary epithelial cells, but are not immortalized cells.关键词
奶牛乳腺上皮细胞/乳房炎/功能性/核型分析Key words
bovine mammary epithelial cells/mastitis/functional/karyotype analysis分类
农业科技引用本文复制引用
詹康,贡笑笑,左晓昕,陈银银,占今舜,赵国琦..奶牛乳腺上皮细胞系的培养与鉴定[J].动物营养学报,2015,(8):2544-2550,7.基金项目
江苏省高校优势学科建设工作资助项目( PAPD) （ PAPD）
