一种提高体外培养细胞中期分裂相的新方法  

A Novel Method for Increasing the Number of Metaphase Cells Cultured in vitro

作　　者：侯颖[1] 谭立新[1] 李文蓉[1] 牛志刚[1] 郭志勤[1] 

机构地区：[1]新疆畜牧科学院农业部家畜繁育生物技术重点开放实验室

出　　处：《遗传》2005年第3期457-460,共4页Hereditas（Beijing)

基　　金：新疆高技术研究发展计划项目。家畜乳腺生物反应器关键技术平台的建立与应用(批准号:200311105)~~

摘　　要：为了提高体外培养细胞中期分裂相,用黄牛胎儿成纤维细胞系(YFF)和西门塔尔小牛成纤维细胞系(CNF)为实验材料,先经4℃低温休克再用秋水仙素处理后制备染色体,比较了不同低温休克处理时间获得的中期分裂相百分率,并运用该方法对20代内YFF和CNF核型变异进行了分析。实验发现,YFF和CNF经低温休克中期分裂相百分率显著高于对照组(P<0.05)。其中,20h低温组中期分裂相(31.7%和40.2%)高于对照组(4.7%和6.4%)5倍以上(P<0.01)。实验结果表明,4℃低温休克法是一种提高体外培养细胞中期分裂相的简便方法,适于监测体外培养细胞核型变异。To increase the number of metaphase cells cultured in vitro, two bovine fibroblast cell lines(YFF and CNF) were frozen at 4℃ for different length of time prior to colchicine treatment, and then chromosomal specimen were prepared. The percentage of metaphase cells was examined under conditions above. Using this method, the variation rate of karyotype of YFF and CNF subcultured up to passage 20 were also analyzed. It was found that the percentage of YFF and CNF metaphase cells in treatment group were significantly higher than that in control group(P<0.05), and the number of YFF and CNF metaphase cells obtained in 20 h treatment group were increased more than 5 fold as many as that in control group(P<0.01), 31.7%and 40.2% vs 4.7%and 6.4%, respectively. These data suggest that the method of freezing at 4℃ could be used for increasing the number of metaphase cells in vitro conveniently, and analyzing the variation rate of karyotype of cultured cells efficiently.

关 键 词：4℃低温休克 中期分裂相 牛成纤维细胞 体外培养 

分 类 号：Q813.1[生物学—生物工程]