新生牛睾丸支持细胞的体外成熟与分化  

In vitro maturation and differentiation of Sertoli cells from neonatal bovine testis

作　　者：郑鹏[1] 冯瑞 秦雪 李琦 赵骞[1] ZHENG Peng;FENG Rui;QIN Xue;LI Qi;ZHAO Qian(College of Animal Science and Technology,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学动物科学技术学院,黑龙江哈尔滨150030

出　　处：《中国兽医学报》2022年第5期981-985,共5页Chinese Journal of Veterinary Science

基　　金：黑龙江省自然科学基金资助项目(LH2020C019)。

摘　　要：为建立支持细胞体外分化成熟的细胞模型,本试验使用犊牛睾丸支持细胞进行传代培养,观察细胞形态的变化,通过免疫荧光染色分析波形蛋白和ZO-1的表达,通过荧光定量RT-PCR和Western blot检测细胞增殖相关基因(PI3K、Akt)、细胞紧密连接相关基因(ZO-1、Connexin-34)的mRNA和蛋白表达的变化。结果表明,随着传代次数增加,支持细胞的细胞质充分扩展,逐渐趋于成熟状态;波形蛋白和ZO-1在未成熟与成熟的支持细胞都呈阳性;支持细胞成熟分化后,PI3K的mRNA和蛋白表达水平显著下降(P<0.05),Akt的mRNA和蛋白表达水平未见明显改变(P>0.05);ZO-1和Connexin-34的mRNA及蛋白表达水平升高,但差异不显著(P>0.05)。体外传代培养能够促进支持细胞的分化成熟。In order to establish a cell model for differentiation and maturation of Sertoli cells in vitro,testicular Sertoli cells were used to subculture for observing the changes in cell morphology.Furthermore,the expression of vimentin and ZO-1 were analyzed by immunofluorescence staining;the changes in mRNA and protein expression of proliferation-related genes(PI3 K,Akt)and cell tight junction-related genes(ZO-1,Connexin-34)were detected by real-time RT-PCR as well as Western blot.The results showed that the cytoplasm of Sertoli cells expanded fully and tended to mature gradually with the increase of passage times;vimentin and ZO-1 were positive in both immature and mature Sertoli cells;mRNA and protein expression of PI3 K significantly decreased(P<0.05)after Sertoli cells maturation and differentiation;no difference was found in mRNA and protein expression of Akt(P>0.05);mRNA and protein expression of ZO-1 and Connexin-34 trended to increase but the difference was not significant as well as(P>0.05).Subculture can promote the differentiation and maturation of Sertoli cells in vitro.

关 键 词：新生牛 睾丸 支持细胞 成熟 分化 

分 类 号：S814[农业科学—畜牧学]