Division of Science and Engineering, Graduate School of Arts and Science, Iwate University, Iwate, 020-8551, Japan.
Institute for Animal Reproduction, Ibaraki, 300-0134, Japan.
Sci Rep. 2023 Mar 3;13(1):3604. doi: 10.1038/s41598-023-30774-x.
Embryo transfer (ET) is an essential reproductive technology for the production of new animal strains and maintenance of genetic resources. We developed a method, named Easy-ET, to induce pseudopregnancy in female rats by artificial stimulation using sonic vibration instead of mating with vasectomized males. This study examined the application of this method for the induction of pseudopregnancy in mice. Offspring were obtained from two-cell embryos transferred into females with pseudopregnancy induced using sonic vibration in proestrus on the day before embryo transfer. Furthermore, high developmental rates of offspring were observed when pronuclear and two-cell embryos were transferred to females in estrus that were stimulated on the day of embryo transfer. Genome-edited mice were also obtained using frozen-warmed pronuclear embryos with clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated system (Cas) nucleases introduced using the technique for animal knockout system by electroporation (TAKE) method, which were transferred to females with pseudopregnancy induced on the day of embryo transfer. This study demonstrated that induction of pseudopregnancy by sonic vibration was also possible in mice.
胚胎移植（ET）是生产新动物品系和维持遗传资源的重要生殖技术。我们开发了一种方法，称为 Easy-ET，通过使用超声波振动对雌性大鼠进行人工刺激来诱导假孕，而不是与输精管切除的雄性交配。本研究探讨了该方法在诱导小鼠假孕中的应用。通过将胚胎转移到发情期的雌性大鼠中，从发情前一天通过超声波振动诱导的假孕雌性大鼠中获得了后代。此外，当将原核和二细胞胚胎转移到发情期当天接受刺激的雌性大鼠中时，观察到后代的高发育率。还使用冷冻解冻的原核胚胎获得了基因组编辑的小鼠，这些胚胎使用电穿孔（TAKE）方法通过引入成簇规律间隔短回文重复（CRISPR）/CRISPR 相关系统（Cas）核酸酶进行编辑，然后将其转移到发情期当天通过超声波振动诱导的假孕雌性大鼠中。本研究表明，超声波振动诱导假孕在小鼠中也是可行的。
