中文摘要：

【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。

英文摘要：

[ Objective ] The aim of this study was to explore the cultural characteristics of testis sertoli cells in Hetian sheep in vitro. [ Method]The experimental testis sertoli cells were obtained from Hetian sheep lamb in 24 h by two-step enzymatic digestion, then the adhesion method was used to purify testis sertoli cells. To observe the testis sertoli cells growth and purity with morphologic method, HE staining detection, AKP staining detection, ORO staining detection, acridine orange staining detection, rhodamine 123 staining detection, immunohistochemical staining detection, RT-PCR, cell counting MTY assay. [ Result ] The results showed that 2. 00 % fetal bovine serum in DMEM/ F-12 medium could be good to culture testis sertoli cells in vitro, the sertoli cells which obtained from Hetian sheep lamb in 24 h could be passaged to 20th generation, the testis sertoli cells contained normal cell morphous and diploid karyotype, the testis sertoli cells expressed vimentin by immunohistoehemical staining, RT-PCR result indicated that the SCF gene and GDNF gene expressed in testis sertoli cells. [ Conclusion ] These results demonstrated that we sueeessed to establish the modle of testis sertoli cells in Hetian sheep.