犊牛睾丸支持细胞的培养及不同浓度pEGFP-N3载体瞬时转染

Culture of testicular Sertoli cells cultured and transient transfection of the pEGFP-N3 vector with the different concentrations in calves

为了研究支持细胞的分离、纯化、鉴定、传代培养及pEGFP-N3载体转染,试验以新生牛睾丸组织为材料,以脂质体介导转染支持细胞。结果表明:支持细胞的纯度达95%,可传至20代;细胞密度为2.5×105个/孔时,3.0μL的脂质体、2.4μg的pEGFP-N3质粒为最佳组合,转染72 h后效率可达36.03%。

To study the separation, purification, identification, subculture of Sertoli cells and the transfection of pEGFP - N3 vector , the testicular tissues of the newborn calves were used as experiment materials, and then the Sertoli cells were transfected with the plasmid pEGFP - N3 by liposome - mediated transfection. The results showed that the purity of the Sertoli ceils reached 95%, and the Sertoli ceils could be passaged to the 20 passages. The best transteetion combination was using 3.0μL of liposomes with 2.4 μg of pEGFP - N3 plasmid when Sertoli cells density was 2.5 × 10^5 ,and the transfeetion efficiency could be reached up to 36.03% at 72 h after transfection.