牛支持细胞和成纤维细胞体外培养及pEGFP-N3转染

Bovine sertoli cells and fibroblast cells culture in vitro and their transfection with the plasmid pEGFP-N3

试验以新生牛睾丸组织、牛胎儿皮肤组织为材料,进行支持细胞、成纤维细胞的分离、纯化、鉴定及pEGFP-N3载体转染。结果表明,完善犊牛支持细胞、胎牛皮肤成纤维细胞体外培养体系,细胞纯度可达95%。支持细胞在相同密度下较成纤维细胞能更快完成细胞生长指数期,生长迅速。转染pEGFP-N3载体后,初期EGFP均匀充满胞浆,后期EGFP向细胞核聚集,荧光强度明显高于细胞质中。转染后24、48、72 h,成纤维细胞转染效率高于支持细胞,转染pEGFP-N3的成纤维细胞和支持细胞在转染48 h后达到最大转染效率。转染后成纤维细胞的存活时间明显高于支持细胞,这与支持细胞生物学性质有关。

The newborn calf testicles and embryo skin were used in this experiment, the sertoli cells and fibroblast cells were transfected with the plasmid pEGFP-N3 by Liposome-mediated. The results showed that the purity of the cells was up to 95%. The early EGFP was evenly filled in cytosol after transfection. Later, EGFP was gathered in the nucleus and the fluorescence intensity in the nucleus became more higher in the cytoplasm. The results showed that transfection efficiency of fibroblast cell was higher than sertoli cells. The transfection efficiency of fibroblast cells and sertoli cells was significantly different after transfection at 24, 48 and 72 h. Both cells got the highest transfection efficiency at 48 h. After transfecting, the life of fibrobLast cells was obviously longer than sertoli cells. It may be due to the inimitable physiological function of sertoli cells and high tansfectiion efficiency of fibroblasts.