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《Heilongjiang Animal Science and Veterinary Medicine》 2012-05
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Identification and in vitro culture of bovine mammary epithelial cells and fibroblast

DING Xu-na,WANG Feng-long,DING Yu-lin,YANG Lei,LI Yu, YIN Jing-feng,ZHAO Zhan-kuan(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China)  
To obtain stabile purified bovine mammary epithelial cells and fibroblast in vitro,the mammary tissue samples were collected from Holstein cows during late gestation or lactation by surgery.After isolated the mammary alveoli,the mammary cells were cultured by tissue explants method in vitro.Then bovine mammary epithelial cells and fibroblasts were purified and cultured separately at different time by trypsin-EDTA digestion and adherent method,and the purity of the cells were identified by immunocytochemical method.The results showed that purified bovine mammary epithelial cells were mainly in polygonal,with round or oval nucleus,the nucleolus were visible and clear,which mainly in cobblestone or paving stone-like growth,and also could secret milk drop,with a positive reaction to cytokeratin-18 and a negative reaction to vimentin,while purified bovine mammary fibroblasts were mainly in spindle,in spiral or radial-shaped growth,with a negative reaction to cytokeratin-18 and a positive reaction to vimentin.After purification,the purity of both cells could exceed ninety-five percent,which could meet the requirement of the follow-up experiments.
【Fund】: 国家自然科学基金项目(30960273)
【CateGory Index】: Q813;S823
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