室温下保存的小鼠屠体卵巢GV卵的发育能力

将ICR系雌性小鼠处死并在10℃、15℃、20℃和25℃下依次保存8、14、24和48 h后,采集其体内的卵巢GV期卵,采用常规方法进行体外成熟和体外受精,获得的2细胞期胚经体外培养或胚胎移植观察其发育能力.其结果,在10℃下保存24 h、15℃下保存14 h、20℃下保存8h和25℃下保存4 h后,其体内附有卵丘细胞的GV卵的体外成熟-体外受精后的2细胞率分别为14%、9%、10%和10%,随着保存温度的提高和保存时间的延长,带有颗粒细胞GV期卵的比率明显降低,同时其GV期卵经体外成熟及体外受精后的2细胞率明显降低.在20℃下保存24 h和25℃下保存14 h时,难以获得形态正常的GV期卵;体外受精获得的2细胞期胚经体外培养,总体上有64%的胚胎发育至扩张囊胚,未见有保存温度和保存时间的显著影响,且利用在15℃保存8 h后的GV卵获得2细胞期胚的移植获得正常新生小鼠.上述结果表明,雌性动物室温条件下死亡后,若能短时间及时采集其体内GV期卵并体外成熟、体外受精,体外培养及胚胎移植技术,就有可能获得新生后代.

Viability of GV Oocytes Collected from Carcasses Kept at Room Temperature

This study was carried out to investigate whether the GV oocytes from carcasses could be utilized. Carcasses were preserved at 10℃, 15℃, 20℃ and 25℃ for 8 hours (as control), 14 hours, 24 hours, or 48 hours after mice were killed and GV occytes were collected. After maturating in vitro, these oocytes were fertilized in vitro with the general procedure in advance. 2-cell embryos were cultured in vitro, then viability was observed. The results showed the fertilizing rate in vitro of the group for GV oocytes with cumulus cells after maturating which were conserved at 10℃ for 24 h, 15℃ for 14 h, 20℃ for 8 h and 25℃ for 4 h was 14%, 9%, 10% and 10% respectively, and with the advance of storage temperature and extension of storage time, the rate of 2-cell after maturating-fertilizating of the GV oocytes decreased significantly. GV oocytes with normal shape were not obtained in the group which was stored at 20℃ for 24 h and at 25℃ for 14 h; A blastula rates (64%) of all groups in average was gotten in culturing 2-cell in vitro, and there is no significant difference in temperature and time of storage. After transplantating some of the 2-cell embryos which were obtained from GV oocytes stored at 15℃ for 8 h, normal infant mice could be obtained. The results above indicated that if we can collect GV oocytes from dead female mice preserved at room temperature in a short time, followed by IVF, IVM, IVC and embryo transplantation, we could obtain new infants.