软琼脂克隆法与裸鼠体内接种法检测细胞致瘤性的比较  

Test for Tumorigenicity of Cells by Soft Agar Cloning Method and Inoculation in Nude Mice

作　　者：孟淑芳[1] 林林[1] 李修兰[1] 冯建平[1] 王佑春[1] 李德富[1] 

机构地区：[1]中国药品生物制品检定所细胞室,北京100050

出　　处：《中国生物制品学杂志》2006年第5期516-519,共4页Chinese Journal of Biologicals

基　　金："自然科技资源共享描述标准整合试点研究与验证完善"项目资助(课题号:2004DKA30360-09)

摘　　要：目的比较体外软琼脂克隆法与裸鼠体内接种法检测细胞致瘤性的敏感性及相关性。方法用软琼脂克隆法检测不同种类、不同来源的细胞的致瘤性,并与裸鼠体内接种法的结果进行比较。结果不同的细胞在体外软琼脂中形成细胞集落的能力及集落形态明显不同。肿瘤细胞在体外可形成肉眼可见的细胞集落,而人二倍体细胞及组织工程用细胞在体外不形成或仅形成极少的细胞集落克隆,其集落形成率与细胞代次没有相关性,在裸鼠体内亦不形成瘤。重组CHO细胞经过遗传改造后,有的在裸鼠体内的致瘤性减弱,但体外软琼脂克隆行为没有发生明显改变。从128至169代次的Vero细胞均可在体外软琼脂中形成不同的细胞集落克隆,高代次细胞的集落形成率明显高于低代次细胞,但所有检测代次的细胞在裸鼠体内均未形成瘤。结论用软琼脂克隆法检测细胞的致瘤性与裸鼠体内接种法不仅具有良好的相关性,而且敏感性高于裸鼠体内接种法。Objective To compare the sensitivities of soft agar cloning method and inoculation in nude mice in test for the tumorigenicity of cells and study the relationship of test results by the two methods. Methods Test the tumorigenicities of cells of various kinds and origins by soft agar cloning method,and compare the result with that by inoculation in nude mice. Results The capacities in forming colony and the morphologies of colonies of various cells in soft agar were significantly different. Tumor cells formed visual colonies in vitro. However,human diploid cells and the tissue engineering cells formed no or few colonies in vitro and formed no tumor in nude mice,and their colony forming rate showed no relationship to the passages. The tumorigenicity of a portion of recombinant CHO cells was attenuated after genetic modification, however, the test result of the cells by soft agar cloning showed no change. The Vero cells of passages 128-169 formed different colonies on soft agar,and the colony forming rate of cells of high passages was significantly higher than that of low passages. However, all the Vero cells of various passages formed no tumor in nude mice. Conclusion The sensitivity of soft agar cloning method was significantly higher than that of inoculation in nude mice in test for the tumorigenicity of ceils, and the test results by the two methods showed good relationship.

关 键 词：软琼脂克隆 体内接种 致瘤性 

分 类 号：R739.41[医药卫生—肿瘤]