In vitro Culture and Characterization of a Mammary Epithelial Cell Line from Chinese Holstein Dairy Cow
The objectivies of this study was to establish a culture system for bovine mammary epithelial cell line in intro, and elucidate the unique characteristics of this cell line. Mammary tissues from a three years old lactating dairy cow (about 100-d) were used as a source of the epithelial cell line, which were cultured in plastic dishes covered with collagen. Fibre cells and epithelial cells successively grew and extended from mammary tissues after cultured for 3d, and pure epithelial cells were obtained by continuous culture and digestion for three times. The strong positive immunocytochemistry to cytokeratin 18 suggested the obtained cell line exhibited the specific character of epithelial cells. The epithelial cells cultured in the presence of 10％ FBS, supraphysiologic concentration of insulin and hydrocortisone maintained a normal diploid chromosome modal number of 2n=60, and were capable of synthesizing and secreting β-casein. The β-casein expression of epithelial cells on molecule and protein level were detected by TR-PCR and Western-bloting methords, respectively. The most important thing is that frozen reservation in 90％FBS and 10％DMSO mixture did not influence the growth character, chromosome number and protein secretion of the obtained epithelial cell line. We conclude that the obtained mammary epithelial cells are normal in morphology, gtowth, cytogenetic and secreted characteristics, and it may become a useful model to study the function of mammary epithelial cells of Chinese Holstein cairy cow (CMECs).
Han Hu Jiaqi Wang Dengpan Bu
State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, China
国际会议
奶牛营养与牛奶质量国际研讨会(ist International Symposium on Dairy Cow Nutrition and Milk Quality)
北京
英文
112-117
2009-05-04（万方平台首次上网日期，不代表论文的发表时间）