绒山羊附睾上皮细胞培养方法的建立及其生长特性

发布时间：2019-05-05 06:26

【摘要】：本研究旨在建立一种简单易行、获取细胞纯度高的绒山羊附睾上皮细胞体外培养体系。10~12月龄绒山羊附睾头部分别采用酶消化法和改良组织块消化法进行原代培养,利用免疫细胞化学染色法和免疫荧光化学法对细胞进行鉴定,同时利用流式细胞仪和CCK法分别检测细胞纯度和增殖情况,在电镜下观察细胞超微结构。结果表明,两种培养方法所获的细胞均呈岛屿状克隆生长和较好的增殖能力,具有活跃的分泌和代谢功能;上皮细胞特异性的角蛋白18和附睾特异性的谷胱甘肽过氧化物酶5(GPx5)的表达以及流式细胞仪结果显示,两种方法所获细胞均为纯度较高的附睾上皮细胞,但改良组织块消化法获得原代细胞需要的时间长,酶消化法获得原代细胞时间短,获得的细胞量大。本研究结果为进一步研究绒山羊附睾上皮功能提供了良好的基础。
[Abstract]:The aim of this study was to establish an in vitro culture system for epididymal epithelial cells of cashmere goats with high cell purity. The epididymal head of 10-12 month-old cashmere goats was primarily cultured by enzyme digestion method and modified tissue block digestion method, respectively. The cell purity and proliferation were detected by flow cytometry and CCK method respectively. The ultrastructure of the cells was observed under electron microscope. The results showed that the cells obtained by the two methods showed island-like clone growth and good proliferation ability, and had active secretory and metabolic functions. The expression of epithelial cell-specific keratin 18 and epididymis-specific glutathione peroxidase 5 (GPx5) and the results of flow cytometry showed that the cells obtained by both methods were high purity epididymal epithelial cells. However, it takes a long time to obtain primary cells by modified tissue block digestion method, and short time to obtain primary cells by enzyme digestion method, and the amount of cells obtained is large. The results of this study provide a good basis for further study of epididymal epithelial function in cashmere goats.
【作者单位】：内蒙古农业大学动物科学学院内蒙古自治区动物遗传育种与繁殖重点实验室;内蒙古医科大学基础医学院;
【基金】：国家自然科学基金(31460598) 海南省重大科技计划(ZDKJ2016017-02)
【分类号】：S827

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