Kawase E, Suemori H, Takahashi N, Okazaki K, Hashimoto K, Nakatsuji N
Division of Developmental Biology, Meiji Institute of Health Science, Odawara, Japan.
Int J Dev Biol. 1994 Jun;38(2):385-90.
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 mutant mouse strains. So far, however, most ES cell lines used in genetic manipulation have been derived from the 129/Sv strain. We have established ES cell lines from blastocysts of the C57BL/6 strain at high efficiency by use of a fibroblast cell line SL10 as feeder cells, and supplementing the culture medium with 5,000 unit/ml LIF. Using such culture conditions, we have also established a number of ES cell lines from the BALB/c, BXSB/MpJ-Yaa and MRL/Mp-lpr/lpr mouse strains. Procedure for establishment of ES cell lines had to be modified among the mouse strains, indicating the strain difference. For example, only a few stem cell colonies appeared from the BALB/c blastocysts. Stem cells of the BXSB strain were very sensitive to the trypsinization in subculturing. Most of these cell lines had normal karyotype and produced chimeric mice. Several C57BL/6 cell lines contributed to the germ-line. These results indicate that ES cell lines can be established from various mouse strains, but their characteristics are different among strains.
为了将小鼠胚胎干细胞(ES细胞)用于各种目的,期望从诸如近交系和突变小鼠品系等各种来源建立细胞系。然而,到目前为止,用于基因操作的大多数ES细胞系都来源于129/Sv品系。我们通过使用成纤维细胞系SL10作为饲养细胞,并在培养基中添加5000单位/毫升白血病抑制因子(LIF),高效地从C57BL/6品系的囊胚中建立了ES细胞系。利用这样的培养条件,我们还从BALB/c、BXSB/MpJ-Yaa和MRL/Mp-lpr/lpr小鼠品系中建立了许多ES细胞系。在不同小鼠品系中,建立ES细胞系的程序必须进行修改,这表明了品系差异。例如,从BALB/c囊胚中仅出现少数干细胞集落。BXSB品系的干细胞在传代培养中对胰蛋白酶消化非常敏感。这些细胞系大多具有正常的核型,并能产生嵌合体小鼠。几个C57BL/6细胞系能形成生殖系。这些结果表明,可以从各种小鼠品系中建立ES细胞系,但它们的特性在不同品系之间存在差异。