品系对小鼠胚胎干细胞分离效率的影响

Effects of Genetic Background on Establishment of Mouse Embryonic Stem Cells

为了充分利用小鼠胚胎干(ES)细胞,就必须从众多小鼠品系中分离ES细胞系。本研究通过传统的成纤维细胞饲养层法,从CD-1、129/Sv、C57BL/6J和129/Sv×C57BL/6J四种不同遗传背景的小鼠中分离得到12个ES细胞系,而从KM小鼠没有得到ES细胞系。所有的ES细胞系都具有典型的ES细胞特征,AKP染色呈阳性。从四种不同遗传背景的ES细胞系得到了包含多种组织的畸胎瘤;与桑椹胚聚合后,都得到了生殖系嵌合体。结果表明:品系对小鼠ES细胞的分离有显著影响,利用129小鼠以及包含129小鼠遗传背景的杂交小鼠都较容易分离ES细胞,由ES细胞得到生殖系嵌合体的效率在不同品系间有显著差异,从杂交ES细胞比近交ES细胞中更容易得到生殖系嵌合体。

For utilization of the mouse embryonic stem (ES) cells for various purposes, it is desirable that the cell lines are established from various sources such as inbred and outbred mouse strains. So far, however, most ES cell lines used in genetic manipulation have been derived from the 129/Sv strain. In this paper, we have established ES cell lines from blastocysts of the CD-1, C57BL/6J and 129/Sv×C57BL/6J mice besides 129/Sv mouse with a fibroblast cell as feeder cells, and supplemented with 1,000 unit/mL LIF in the culture medium. Twelve ES cell lines were obtained from CD-1, 129/Sv, C57BL/6J and 129/Sv×C57BL/6J mouse blastocysts. No ES cell line from KM embryos suggests that KM strain might have inhibitory genetic factor(s) for the ES cell formation. Some ES cell lines could be obtained from hybrids at high efficiency suggested a heterosis effect can be expected for establishing ES cell lines in mice. These ES cell lines produced chimeric mice and contributed to the germ-line. The results indicate that genetic background is an important factor in isolating ES cells.