Park Min-Jee, Kim Eun-Young, Kang Man-Jong, Lee Jun-Beom, Jeong Chang-Jin, Park Se-Pill
1 Jeju National University Stem Cell Research Center , Seoul, Korea.
2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Korea.
Cell Reprogram. 2017 Aug;19(4):245-254. doi: 10.1089/cell.2017.0003. Epub 2017 Jun 26.
A time-lapse monitoring system has predictive value for selecting good-quality embryos with the highest implantation potential. Using this new tool, we investigated the developmental potential and developmental kinetics of bovine parthenogenetic (PA) and two types of somatic cell nuclear transfer (NT) embryos. Bovine non-transgenic ear cells (bECs) or transgenic cells (bTGCs) were used as donor cells. The cleavage and blastocyst development rates did not significantly differ among the PA, NT-bEC, and NT-bTGC groups, and first cleavage occurred an average of 19.3 hours (n = 70), 21.6 hours (n = 60), and 21.3 hours (n = 62) after activation, respectively (20.4 hours [n = 192] for all embryos). When embryos were classified into early cleaving (≤20 hours) and late cleaving (>20 hours) groups, the blastocyst formation rate was much higher in the early cleaving groups (PA, 46%; NT-bEC, 50%; NT-bTGC, 39%) than in the late cleaving groups (PA, 18%; NT-bEC, 23%; NT-bTGC, 28%), while the percentage of embryos whose development was blocked between the two- and eight-cell stages was increased in the late cleaving groups. The percentage of embryos classified as early cleaving with a normal morphology was twofold higher in the PA group (20.0%, n = 14) than in the NT-bTGC group (9.7%, n = 6). The timing of each developmental stage varied widely; the timing of first cleavage varied from 7.6 hours in the PA group to 34.5 hours in the NT-bEC group and the timing of expanded/hatching blastocyst appearance varied from 141.6 hours in the PA group to 196.3 hours in the NT-bTGC group, differences of 26.9 and 54.7 hours, respectively (PA>NT-bEC>NT-bTGC). These results demonstrate that time-lapse monitoring provides novel data regarding individual embryo developmental kinetics and helps to predict developmental potential for improved bovine NT embryo selection based on early cleavage (≤20 hours) and normal morphology.
延时监测系统对于选择具有最高着床潜力的优质胚胎具有预测价值。利用这一新工具,我们研究了牛孤雌生殖(PA)胚胎和两种类型的体细胞核移植(NT)胚胎的发育潜力和发育动力学。牛非转基因耳细胞(bECs)或转基因细胞(bTGCs)用作供体细胞。PA组、NT-bEC组和NT-bTGC组之间的卵裂率和囊胚发育率没有显著差异,首次卵裂分别发生在激活后平均19.3小时(n = 70)、21.6小时(n = 60)和21.3小时(n = 62)(所有胚胎为20.4小时[n = 192])。当胚胎被分为早期卵裂(≤20小时)组和晚期卵裂(>20小时)组时,早期卵裂组的囊胚形成率(PA组为46%;NT-bEC组为50%;NT-bTGC组为39%)远高于晚期卵裂组(PA组为18%;NT-bEC组为23%;NT-bTGC组为28%),而晚期卵裂组中发育阻滞在二细胞和八细胞阶段之间的胚胎百分比增加。形态正常且被归类为早期卵裂的胚胎百分比在PA组(20.0%,n = 14)中是NT-bTGC组(9.7%,n = 6)的两倍。每个发育阶段的时间差异很大;首次卵裂时间从PA组的7.6小时到NT-bEC组的34.5小时不等,扩张/孵化囊胚出现的时间从PA组的141.6小时到NT-bTGC组的196.3小时不等,分别相差26.9小时和54.7小时(PA>NT-bEC>NT-bTGC)。这些结果表明,延时监测提供了关于单个胚胎发育动力学的新数据,并有助于基于早期卵裂(≤20小时)和正常形态来预测发育潜力,以改进牛NT胚胎的选择。