大鼠附睾上皮细胞的原代培养及纯化鉴定  

Purification and characterization of primary cultured rat epididymal epithelial cells

作　　者：程胖[1] 刘博[1] 冯潇[1] 魏金花[1] 马斌芳[1] 李臻[1] Cheng Pang;Liu Bo;Feng Xiao;Wei Jinhua;Ma Binfang;Li Zhen(Department of Human Anatomy,Histology and Embryology,the Fourth Military Medical University,XVan 710032,China)

机构地区：[1]第四军医大学人体解剖与组织胚胎学教研室,西安710032

出　　处：《中国组织化学与细胞化学杂志》2018年第3期232-236,共5页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金(31271606);陕西省自然科学基金(2015JM8403)

摘　　要：目的建立大鼠附睾上皮细胞原代培养及纯化方法。方法利用酶消化法和组织块法对大鼠附睾上皮细胞进行原代培养,然后用胰酶两步消化法进一步纯化附睾上皮细胞,最后分别利用免疫荧光和免疫组织化学染色对原代培养的细胞及相关蛋白表达情况进行鉴定。结果酶消化法较组织块法得到的附睾上皮细胞纯度高,免疫荧光染色结果证明所得附睾上皮细胞主要是主细胞,免疫组织化学结果证明培养的附睾上皮细胞中有雄激素受体和雌激素受体α的表达。结论利用酶消化法对大鼠附睾上皮细胞进行体外培养,方法简单易行,成功率高。Objective To establish汪dissociation and purification method to culture prim ary rat epididymal epithelial cells m vitro.Methods Primary rat epididymal epithelial cells were isolated using either tissue explant or enzyme digestion.The cells in culture were farther purified through two-step trypsin digestion.Finally,the purity of prim ary epididymal epithelial cell culture was assessed by immunocytochemistry(ICC)and immunohistochemistry using clusterin,androgen receptor(AR)and estrogen receptor a(ERa)as markers.Results ICC results indicated that the rat epididymal epithelial cells isolated through enzyme digestion had a higher percentage of principal cells in the culture than through tissue explant.Immunohistochemistry confirmed the expression of both androgen receptor(AR)and estrogen receptor a(ERa)in the prim ary epididymal epithelial cells.Conclusion Isolating rat epididymal epithelial cells through enzyme digestion method is simple and easy w ith higher success rate.

关 键 词：附睾上皮细胞 原代培养 纯化 雄激素受体 雌激素受体a 

分 类 号：Q2-33[生物学—细胞生物学]