CAJ | 학술논문

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.
장ICR계자성소서처사병재10℃、15℃、20℃화25℃하의차보존8、14、24화48 h후,채집기체내적란소GV기란,채용상규방법진행체외성숙화체외수정,획득적2세포기배경체외배양혹배태이식관찰기발육능력.기결과,재10℃하보존24 h、15℃하보존14 h、20℃하보존8h화25℃하보존4 h후,기체내부유란구세포적GV란적체외성숙-체외수정후적2세포솔분별위14%、9%、10%화10%,수착보존온도적제고화보존시간적연장,대유과립세포GV기란적비솔명현강저,동시기GV기란경체외성숙급체외수정후적2세포솔명현강저.재20℃하보존24 h화25℃하보존14 h시,난이획득형태정상적GV기란;체외수정획득적2세포기배경체외배양,총체상유64%적배태발육지확장낭배,미견유보존온도화보존시간적현저영향,차이용재15℃보존8 h후적GV란획득2세포기배적이식획득정상신생소서.상술결과표명,자성동물실온조건하사망후,약능단시간급시채집기체내GV기란병체외성숙、체외수정,체외배양급배태이식기술,취유가능획득신생후대.
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.