山羊耳皮肤成纤维细胞的传代培养及活力分析

目的　研究体外传代培养山羊耳皮肤成纤维细胞活力及染色体倍性。　方法　体外培养山羊耳皮肤成纤维细胞至 17代 ,并利用TUNEL原位分析、BrDU结合免疫标记以及低渗悬滴制备染色体的方法 ,对第 5代和第 17代细胞的细胞凋亡、DNA合成和染色体倍性进行了分析。　结果　山羊皮肤成纤维细胞在体外能够传代到17代以上 ,85 %以上 (42 4 9)的细胞染色体为正常的 2倍体 ,并且与第 5代的细胞相比较 ,细胞凋亡 (1 75 %vs1 15 % )、DNA合成 (17 4 3%vs 16 89% )都没有显著的变化 (P >0 0 5 )。　结论　长期传代培养 (17代 )的成纤维细胞的活力和DNA物质没有受到严重损伤 ,可以用来作为核移植的供体

In Vitro CULTURE AND VIABILITY ANALYSIS OF FIBROBLASTS FROM THE GOAT EAR SKIN

Objective Fibroblasts from goat ear skin were subcultured,cell viability and chromosome ploidy were examined to provide a large number of donor cells for somatic nuclear transfer. Methods Fibroblasts from goat ear skin were cultured in vitro, and cell apoptosis,DNA synthesis and chromosome ploidy of 15th passage and 17th passage cells were estimated by TUNEL,BrDU incorporation and chromosome analysis. Results Above 85%(42/49) cells cultured to 17th passage had normal diploid chromosomes,and the percentages of cells undergoing appoptosis(1 75% vs 1 15%)or DNA synthesis(17 43% vs 16 89%)didn't change significantly compared with 5th passage cells.Conclusion No singificant damage was found in viability and chromosome ploidy of cells cultured up to 17th passage,and fibroblasts from goat ear skin cultured in vitro for a long time could be used as nuclear transfer donors.