Establishment and chimera analysis of 129/SvEv- and C57BL/6-derived mouse embryonic stem cell lines
Establishment and chimera analysis of 129/SvEv- and C57BL/6-derived mouse embryonic stem cell lines
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DOI：
10.2144/00295st04
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发表时间：
2000-11-01
期刊：
影响因子：
2.7
通讯作者：
Joyner, AL
中科院分区：
文献类型：
--
作者：
Auerbach, W;Dunmore, JH;Joyner, AL
每年都有数百种新的突变小鼠品系在胚胎干细胞（ES）中使用基因靶向和基因陷阱方法产生，随着人类和小鼠基因组计划的进展，这一数字预计将继续增长。强大的ES细胞系和简单的嵌合体制造技术的可用性现在比以前更有吸引力。我们从129/SvEv和C57 BL/6小鼠中建立了几种新的ES细胞系，并测试了它们在囊胚注射和/或更便宜和更容易的桑椹胚-ES细胞聚集方法后对种系的贡献能力。使用桑椹胚聚集产生嵌合体，发现小鼠新衍生的129/SvEx和在早期传代时测试的两个C57 BL/6 ES细胞系广泛地促进嵌合体并产生生殖系传递雄性嵌合体。此外，所测试的两个129/vEv ES细胞系和其中一个C57 BL/6 ES细胞系能够在体外多次传代后保持这些特征。我们的研究结果表明，胚胎干细胞的能力，有助于强烈的嵌合体聚集后与远系繁殖的胚胎是早期传代胚胎干细胞的一般属性，并可以保持许多代。然而，C56 BL/6衍生的ES细胞系比129衍生的ES细胞系具有更大的倾向，失去它们在生殖系中定殖的能力。
Hundreds of new mutant mouse lines are being produced annually using gene targeting and gene trap approaches in embryonic stem (ES) cells, and the number is expected to continue to grow as the human and mouse genome projects progress. The availability of robust ES cell lines and a simple technology for making chimeras is more attractive now than over before. We established several new ES cell lines from 129/SvEv and C57BL/6 mice and tested their ability to contribute to the germline following blastocyst injections and/or the less expensive and easier method of morula-ES cell aggregation. Using morula aggregation to produce chimeras, mice newly derived 129/SvEx and two C57BL/6 ES cell lines tested at early passages were found to contribute extensively to chimeras and produce germline-transmitting male chimeras. Furthermore, the two 129/vEv ES cell lines that were tested and one of the C57BL/6 ES cell lines were able to maintain these characterisatics after many passages in vitro. Our results indicate that the ability of ES cells to contribute strongly to chimeras following aggregation with outbred embryos is a general property of early passage ES cells and can be maintained for many passages. C56BL/6-derived ES cell lines, however, have a greater tendancy than 129-derived ES cell lines to lose their ability to colonize the germline.