中文摘要：

本研究以荷斯坦奶牛乳腺为研究对象,旨在建立奶牛乳腺干细胞体外分离培养方法及纯化体系并对其进行初步鉴定。消毒清洗后的组织小块400 IU/m L胶原酶、37℃环境下消化20 h,初步分离得到乳腺干细胞,悬滴培养法对其进行筛选纯化,再对纯化乳腺干细胞的候选表面标记进行检测。结果表明：获得的干细胞具有自我增殖能力,并在悬滴培养条件下能够形成类似乳腺球结构细胞团,而且,不仅表达干细胞标记基因Sox2、Nanog,还表达乳腺干细胞参考标记基因CD24、CD29、CD49f,初步认定分离纯化的细胞为乳腺干细胞。

英文摘要：

This research was designed to establish the separation of holstein cow m ammary stem cells in vitro culture method and purification system.By using 400 IU/m L collagenase digestion,37 ℃ environment 20 h,preliminary mammary stem cells were isolated from mammary tissue.Then through hanging drop culture method to screen mammary stem cells obtained by preliminary separation purification,then purification of cells after gene level verification.The results showed that the stem cells with self proliferation and under the condition of hanging drop culture can form the structure similar to mammospheres.Mammary stem cells can not only express the stem cell marker genes,but also express the reference marker genes of mammary stem cell.Preliminary determination of cells after purification for mammary stem cells.