高效建立129/ter、C57BL/6J小鼠胚胎干细胞系的方法学探讨  

Discussion of the Methods for Establishing Embryonic Stem Cell Lines from 129/ter、C57BL/6J Mice with High Efficiency

作　　者：孟国良[1] 汤富酬[1] 尚克刚[1] 薛友纺[1] 

机构地区：[1]北京大学生命科学学院遗传教研室,北京100871

出　　处：《生物工程学报》2002年第6期740-743,共4页Chinese Journal of Biotechnology

基　　金：国家自然科学基金资助项目 (No .30 0 70 414 )~~

摘　　要：A new method for establishing ES cell lines from 129/ter、C57BL/6J mice was set up which was characterized by the murine embryonic fibroblast cell(MEF) feeder, the medium of rat heart cell-conditioned medium(RH-CM)for ES cells, and the consecutive digestion by the digestion liquid containing 1% serum. Every group of improved experiments was done with a control of routine method. The results showed that, compared with routine method, the improved way increased the ratio of ES cell lines of 129/ter mice from 11.8% to 33.3%, and of C57BL/6J from 3.7% to 13.3%. The difference is distinct. The passage culture of ES cells showed that, compared with medium added LIF, RH-CM not only inhibited the differentiation of murine ES cells, maintained its dipoild karyotype, but also promote its adherence growth. This kind of culture condition not only maintained the ES cells in an undifferentiated state and their normal dipoild karyotype, but also a series of other characteristics of totipotent embryonic stem cells during extended culture period.A new method for establishing ES cell lines from 129/ter、C57BL/6J mice was set up which was characterized by the murine embryonic fibroblast cell(MEF) feeder, the medium of rat heart cell-conditioned medium(RH-CM)for ES cells, and the consecutive digestion by the digestion liquid containing 1% serum. Every group of improved experiments was done with a control of routine method. The results showed that, compared with routine method, the improved way increased the ratio of ES cell lines of 129/ter mice from 11.8% to 33.3%, and of C57BL/6J from 3.7% to 13.3%. The difference is distinct. The passage culture of ES cells showed that, compared with medium added LIF, RH-CM not only inhibited the differentiation of murine ES cells, maintained its dipoild karyotype, but also promote its adherence growth. This kind of culture condition not only maintained the ES cells in an undifferentiated state and their normal dipoild karyotype, but also a series of other characteristics of totipotent embryonic stem cells during extended culture period.

关 键 词：129/ter小鼠 C57BL/6J小鼠 胚胎干细胞系 方法学 大鼠心脏细胞条件培养 建系 二倍体核型 

分 类 号：Q813.1[生物学—生物工程]