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奶山羊乳腺上皮细胞系的建立
引用本文:佟慧丽,尹德云,李庆章,高学军.奶山羊乳腺上皮细胞系的建立[J].东北农业大学学报,2008,39(8).
作者姓名:佟慧丽  尹德云  李庆章  高学军
作者单位:东北农业大学乳品科学教育部重点实验室,哈尔滨,150030
基金项目:东北农业大学创新团队发展计划
摘    要:乳腺上皮细胞系的建立为乳蛋白基因表达调控机制的研究及乳腺表达载体的检测提供有利条件。本研究建立了正常培养的奶山羊乳腺上皮细胞系。利用胶原酶和透明质酸消化法从奶山羊乳腺组织中分离并获得了纯化的乳腺上皮细胞,利用细胞角蛋白18的免疫荧光染色对乳腺上皮细胞进行了鉴定。通过细胞生长曲线、群体倍增时间、细胞接种存活率、细胞活力检测等生物学性状的检测建立并鉴定了奶山羊乳腺上皮细胞系。结果表明,奶山羊乳腺上皮细胞在添加表皮生长因子、胰岛素样生长因子-1、转铁蛋白-硒钠、10%胎牛血清的DF12培养液中生长状态良好,细胞传至30代时仍保持旺盛的增殖活力。通过细胞传代及反复冻存和复苏实现了细胞系的长期保存,建立了乳腺上皮细胞系,获得了大量的乳腺上皮细胞。从而为开展奶山羊、奶牛等产奶动物乳腺生物反应器及泌乳机制的研究提供了便利条件。

关 键 词:奶山羊  乳腺上皮细胞  细胞系

Establishment of dairy goat mammary gland epithelial cell line
TONG Huili,YIN Deyun,LI Qingzhang,GAO Xuejun.Establishment of dairy goat mammary gland epithelial cell line[J].Journal of Northeast Agricultural University,2008,39(8).
Authors:TONG Huili  YIN Deyun  LI Qingzhang  GAO Xuejun
Abstract:Establishment of mammary gland cell lines is convenient for the study of milk protein gene expression regulation and mammary gland expression vector detection.A normal cultured dairy goat mammary gland epithelial cell line was established in this research.Purified mammary gland epithelial cells were obtained from dairy goat by digestion with collagen enzyme and hyaluronidase,and they were verified by immunofluo-rescent staining of cytokeratin 18.Cell growth curve,doubling time of cell population,cell viability and other cell biological characters were detected.The results showed that mammary gland epithelial cells kept good growth state in culture fluid DF12 with addition of EGF,IGF-1,ITS and 10% FBS.Cells having passed 30 generations still remained good proliferation vital force.Cell passage and resuscitation could make mammary gland epithelial cell line conserve for a long time,a mammary epithelial cell line was established and a great quantity of mammary gland epithelial cells were obtained.The result is convenient for the study of mammary gland bioreactor and lactation mechanism of dairy goats and dairy cows and other economic animals.
Keywords:dairy goat  mammary gland epithelial cell  cell line
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