和田羊睾丸支持细胞的体外培养与分离鉴定

发布时间：2018-05-29 23:32

本文选题：和田羊 + 睾丸　；参考：《西南农业学报》2017年08期

【摘要】：【目的】研究和田羊睾丸支持细胞的体外培养特性。【方法】取新生和田羊睾丸组织,利用两步酶消化后,采用差速贴壁法纯化细胞进行体外培养,并通过形态学观察、HE染色、AKP染色、油红O染色、吖啶橙染色、罗丹明123染色、免疫组化染色、RTPCR和MTT法检测细胞活性及纯度。【结果】培养基中加入2.00%血清浓度的DMEM/F-12可提高支持细胞纯度,在体外培养传至20代的和田羊睾丸支持细胞形态和核型均为正常;所培养的细胞内波形蛋白表达及标志基因SCF和GDNF的表达均为阳性。【结论】本试验成功建立了和田羊睾丸支持细胞体外培养模型。
[Abstract]:[Objective] to study the in vitro culture of sheep testis support cells. [Methods] the cells were cultured in vitro by two step enzyme digestion, and the cells were cultured in vitro by two step enzymes. The cells were stained by morphological observation, staining with AKP, staining with oil red O, staining with acridine orange, Luo Danming 123 staining, immunohistochemical staining, RTPCR and MTT methods were used to detect cell activity and purity. [results] the addition of 2% serum concentration of DMEM/F-12 in the medium could improve the purity of the support cells. The morphology and karyotype of the testis support cells in the 20 generations of the cultured sheep were normal, and the expression of vimentin and the marker gene SCF and GDNF in the cultured cells were all positive. [Conclusion] the model of Sertoli cell culture in Hetian Sheep was successfully established in this experiment.
【作者单位】：塔里木大学动物科学学院/新疆生产建设兵团塔里木畜牧科技重点实验室;
【基金】：国家自然科学基金(31460678) 国家星火计划项目(2015GA891015) 兵团塔里木畜牧科技重点实验室开放课题(HS201409)
【分类号】：S826

【相似文献】