Ottawa Hospital Research Institute, 501 Smyth Road, Ottawa, ON, K1H 8L6, Canada.
Transgenic Res. 2010 Dec;19(6):1137-44. doi: 10.1007/s11248-010-9369-6. Epub 2010 Feb 9.
Generation of mouse chimeras is useful for the elucidation of gene function. In the present report, we describe a new technique for the production of chimeras by injection of R1 embryonic stem (ES) cells into the perivitelline space of one-cell stage mouse embryos. One-cell embryos are injected with 2-6 ES cells into the perivitelline space under the zona pellucida without laser-assistance. Our embryo culture experiments reveal that ES cells injected at the one-cell stage embryo start to be incorporated into the blastomeres beginning at the 8-cell stage and form a chimeric blastocyst after 4 days. We have used this approach to successfully produce a high rate of mouse chimeras in two different mouse genetic backgrounds permitting the establishment of germ line transmitters. This method allows for the earlier introduction of ES cells into mouse embryos, and should free up the possibility of using frozen one-cell embryos for this purpose.
制备嵌合体小鼠对于阐明基因功能非常有用。在本报告中，我们描述了一种通过将 R1 胚胎干细胞（ES 细胞）注入单细胞期小鼠胚胎的卵周隙来制备嵌合体的新技术。在没有激光辅助的情况下，将 2-6 个 ES 细胞注入透明带下的卵周隙中。我们的胚胎培养实验表明，在单细胞期胚胎中注射的 ES 细胞从 8 细胞期开始被整合到卵裂球中，并在 4 天后形成嵌合囊胚。我们已经使用这种方法在两种不同的小鼠遗传背景下成功地产生了高比例的嵌合体小鼠，从而建立了种系传递者。这种方法允许更早地将 ES 细胞引入小鼠胚胎中，并可能为使用冷冻的单细胞胚胎提供可能性。
