Takeo Toru, Kondo Tomoko, Haruguchi Yukie, Fukumoto Kiyoko, Nakagawa Yoshiko, Takeshita Yumi, Nakamuta Yuko, Tsuchiyama Shuuji, Shimizu Norihiko, Hasegawa Takanori, Goto Motohito, Miyachi Hitoshi, Anzai Masayuki, Fujikawa Rie, Nomaru Koji, Kaneko Takehito, Itagaki Yoshiaki, Nakagata Naomi
Division of Reproductive Engineering, Center for Animal Resources and Development (CARD), Kumamoto University, Kumamoto, Japan.
J Am Assoc Lab Anim Sci. 2010 Jul;49(4):415-9.
At refrigerated temperatures, mouse embryos can maintain developmental ability for short periods. Previously, we succeeded in transporting vitrified and warmed 2-cell mouse embryos while maintaining developmental ability at refrigerated temperatures for 50 h. Transport of nonfrozen embryos is an easier and more useful means of exchanging genetically engineered mice between laboratories than is transport of cryopreserved embryos. Here we examined the developmental ability of transported 2-cell embryos that were produced through in vitro fertilization using cryopreserved sperm. Results show that 2-cell embryos produced by cryopreserved sperm can develop into blastocysts after cold storage for 24, 48, and 72 h. Transported 2-cell embryos produced by cryopreserved sperm yielded a favorable number of pups in all of the receiving laboratories after transport lasting 48 to 52 h. In summary, cold storage and transport of 2-cell embryos derived from cryopreserved sperm at refrigerated temperatures provides a novel means of transporting genetically engineered mice as an alternative to the transport of cryopreserved embryos and sperm.
在冷藏温度下,小鼠胚胎能够在短时间内维持发育能力。此前,我们成功运输了玻璃化并复温的2细胞期小鼠胚胎,同时在冷藏温度下将其发育能力维持了50小时。与运输冷冻保存的胚胎相比,运输未冷冻的胚胎是在实验室之间交换基因工程小鼠的一种更简便且更实用的方式。在此,我们研究了使用冷冻保存的精子通过体外受精产生的运输2细胞期胚胎的发育能力。结果表明,由冷冻保存的精子产生的2细胞期胚胎在冷藏24、48和72小时后能够发育成囊胚。在持续48至52小时的运输后,由冷冻保存的精子产生的运输2细胞期胚胎在所有接收实验室都产出了数量可观的幼崽。总之,在冷藏温度下对源自冷冻保存精子的2细胞期胚胎进行冷藏和运输,为运输基因工程小鼠提供了一种新方法,可替代运输冷冻保存的胚胎和精子。