奶牛耳成纤维细胞的分离培养及培养条件的研究  

Isolation and Purification of Cattle Ear Fibroblast and Research on Culture Conditions

作　　者：霍道坦[1] 陶勇[1] 田宁宁[1] 权青[1] 白海[1] 胡敏兰[1] 张运海[1] 章孝荣[1] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036

出　　处：《中国奶牛》2008年第6期14-17,共4页China Dairy Cattle

基　　金：安徽省"十一五"科技攻关课题(04003005);安徽省优秀青年科技基金(06041081)

摘　　要：本试验研究了奶牛耳成纤维细胞的体外培养,培养液、消化液、胰岛素和血清饥饿对细胞的影响。采用组织块贴壁法培养了奶牛耳成纤维细胞,采用0.25%胰蛋白酶差别消化获得了纯化的奶牛耳成纤维细胞。用三种不同的培养液即DMEM(低糖)、DMEM(高糖)、DMEM/F12,对细胞进行培养,结果细胞群体倍增时间均在2.3～2.5d,DMEM(高糖)的培养效果稍好。用三种不同的消化液(0.25%胰蛋白酶,0.02%EDTA,0.25%胰蛋白酶+0.02% EDTA)对传代细胞进行消化,结果表明0.25%胰蛋白酶+0.02%EDTA所用时间短,消化后培养细胞贴壁率高。在培养液中添加胰岛素能促进细胞的生长。对指数生长期的细胞进行血清饥饿后,细胞仍保持着较高的活力。This experiment studied the culture of cattle ear fibroblast in vitro, including medium, digestion, insulin and serum starvation. Adopting the organization piece stuck a wall method to culture the cattle ear fibroblast, adopted 0.25% trypsin in difference time to digest the cattle ear fibroblast. Use three kinds of different medium cultivate the cells,the result shows that DMEM（high sugar） is slightly good. Use three kinds of different digestive（0.25% trysin,0.02% EDTA, 0.25% trypsin＋0.02% EDTA） to digest ear fibroblast cell, the result indicates the time of 0.25% trypsin＋0.02% EDTA digesting group is shortest and the rate of adherence is high after digest. The medium of insulin can promote the growth of ear fibroblast. Performing the serum starvation cultivation to the cells in logarithmic growth Phase,the cell still have high vital force.

关 键 词：耳成纤维细胞 体外培养 血清饥饿 髓岛素 奶牛 

分 类 号：S823[农业科学—畜牧学]