Efficient production of mice from embryonic stem cells injected into four- or eight-cell embryos by piezo micromanipulation
Efficient production of mice from embryonic stem cells injected into four- or eight-cell embryos by piezo micromanipulation
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DOI：
10.1634/stemcells.2008-0164
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发表时间：
2008-01-01
期刊：
影响因子：
5.2
通讯作者：
Liu, Lin
中科院分区：
文献类型：
--
作者：
Huang, Junjiu;Deng, Kai;Liu, Lin
生产胚胎干细胞衍生的基因敲除或转基因小鼠的传统方法是将胚胎干细胞注射到正常的二倍体囊胚中，然后进行几轮嵌合体的繁殖，因此是一个耗时且低效的过程。F0胚胎幼崽也可以直接从四倍体胚胎互补中获得，这需要两细胞胚胎的融合。最近，利用激光辅助微操作系统将胚胎干细胞注射到8细胞胚胎中，产生了F0胚胎幼崽。我们报告了一种简单的方法，通过将胚胎干细胞压电注射到四细胞或八细胞胚胎中，产生F0胚胎生殖系胜任小鼠。用这种方法生产活的转基因小鼠的效率比用四倍体囊胚互补法高。这种直接产生F0胚胎干细胞后代的高效和经济的技术可以应用于许多实验室中，使用胚胎干细胞技术创造基因操纵小鼠，也可以用于严格测试新的胚胎干细胞或其他类型的多能干细胞系的发育能力。
The conventional method for producing embryonic stem (ES) cell-derived knockout or transgenic mice involves injection of ES cells into normal, diploid blastocysts followed by several rounds of breeding of resultant chimeras and thus is a time-consuming and inefficient procedure. F0 ES cell pups can also be derived directly from tetraploid embryo complementation, which requires fusion of two-cell embryos. Recently, F0 ES cell pups have been produced by injection of ES cells into eight-cell embryos using a laser-assisted micromanipulation system. We report a simple method for producing F0 ES cell germline-competent mice by piezo injection of ES cells into four- or eight-cell embryos. The efficiency of producing live, transgenic mice by this method is higher than that with the tetraploid blastocyst complementation method. This efficient and economical technique for directly producing F0 ES cell offspring can be applicable in many laboratories for creating genetically manipulated mice using ES cell technology and also for stringent testing of the developmental potency of new ES cell or other types of pluripotent stem cell lines.