Tokoro Mikiko, Fukunaga Noritaka, Yamanaka Kaori, Itoi Fumiaki, Terashita Yukari, Kamada Yuko, Wakayama Sayaka, Asada Yoshimasa, Wakayama Teruhiko
Asada Institute for Reproductive Medicine, Asada Ladies Clinic Medical Corporation, Kasugai, Aichi, Japan.
Laboratory for Mouse Genetic Engineering, Quantitative Biology Center RIKEN, Suita, Osaka, Japan.
PLoS One. 2015 Sep 22;10(9):e0138854. doi: 10.1371/journal.pone.0138854. eCollection 2015.
Generally, transportation of preimplantation embryos without freezing requires incubators that can maintain an optimal culture environment with a suitable gas phase, temperature, and humidity. Such incubators are expensive to transport. We reported previously that normal offspring were obtained when the gas phase and temperature could be maintained during transportation. However, that system used plastic dishes for embryo culture and is unsuitable for long-distance transport of live embryos. Here, we developed a simple low-cost embryo transportation system. Instead of plastic dishes, several types of microtubes-usually used for molecular analysis-were tested for embryo culture. When they were washed and attached to a gas-permeable film, the rate of embryo development from the 1-cell to blastocyst stage was more than 90%. The quality of these blastocysts and the rate of full-term development after embryo transfer to recipient female mice were similar to those of a dish-cultured control group. Next, we developed a small warm box powered by a battery instead of mains power, which could maintain an optimal temperature for embryo development during transport. When 1-cell embryos derived from BDF1, C57BL/6, C3H/He and ICR mouse strains were transported by a parcel-delivery service over 3 days using microtubes and the box, they developed to blastocysts with rates similar to controls. After the embryos had been transferred into recipient female mice, healthy offspring were obtained without any losses except for the C3H/He strain. Thus, transport of mouse embryos is possible using this very simple method, which might prove useful in the field of reproductive medicine.
一般来说,运输未冷冻的植入前胚胎需要能通过合适的气相、温度和湿度维持最佳培养环境的培养箱。这种培养箱运输成本高昂。我们之前报道过,在运输过程中若能维持气相和温度,就能获得正常后代。然而,该系统使用塑料培养皿进行胚胎培养,不适用于活体胚胎的长途运输。在此,我们开发了一种简单低成本的胚胎运输系统。我们测试了几种通常用于分子分析的微管,以替代塑料培养皿用于胚胎培养。当对其进行清洗并附着在透气膜上时,从1细胞到囊胚阶段的胚胎发育率超过90%。这些囊胚的质量以及将胚胎移植到受体雌性小鼠后足月发育的比率与培养皿培养的对照组相似。接下来,我们开发了一种由电池而非市电供电的小型保温箱,它能在运输过程中维持胚胎发育的最佳温度。当使用微管和保温箱通过包裹递送服务对来自BDF1、C57BL/6、C3H/He和ICR小鼠品系的1细胞胚胎进行3天运输时,它们发育成囊胚的比率与对照组相似。在将胚胎移植到受体雌性小鼠后,除C3H/He品系外,均获得了健康后代且无任何损失。因此,使用这种非常简单的方法运输小鼠胚胎是可行的,这在生殖医学领域可能会被证明是有用的。