Ideta Atsushi, Urakawa Manami, Aoyagi Yoshito, Saeki Kazuhiro
Zen-noh Embryo Transfer Center, Kamishihoro, Katogun, Hokkaido, Japan.
Cloning Stem Cells. 2007 Winter;9(4):571-80. doi: 10.1089/clo.2007.0017.
Bovine somatic cell nuclear transfer (NT) embryos can develop to normal calves, but the success rates are still quite low. Recently, enhanced development of bovine NT embryos to full term has been achieved using fibroblasts at the early G1 phase instead of cells at the quiescent (G0) phase. In the present study, we examined the morphological development in utero of NT embryos using early G1 phase cells (eG1-NT embryos) and G0 phase cells (G0-NT embryos). We produced eG1- and G0-NT blastocysts, and then they were transferred to recipient heifers for transient development in utero up to day 14 of gestation. In vitro-fertilized (IVF), parthenogenetic and artificially inseminated (AI) embryos were used as controls. The rate of formation of embryonic disks of the recovered embryos was the same among the groups of eG1-NT, IVF, and AI embryos (p>0.05). The formation rate in eG1-NT embryos was significantly higher than that in G0-NT embryos (p<0.05). The lengths of eG1-NT embryos were the same as those of IVF, parthenogenetic, and AI embryos (p>0.05), but significantly shorter than those of G0-NT embryos (p<0.01). We conclude that the morphological development of day 14 embryos derived from eG1-NT embryos was mostly similar to that of AI embryos, but that the morphological development of G0-NT embryos was abnormally large and different from that of AI and eG1-NT embryos.
牛体细胞核移植(NT)胚胎能够发育成正常的犊牛,但成功率仍然相当低。最近,通过使用处于G1早期阶段的成纤维细胞而非静止(G0)期的细胞,已实现牛NT胚胎发育至足月。在本研究中,我们检查了使用G1早期阶段细胞(eG1-NT胚胎)和G0期细胞(G0-NT胚胎)的NT胚胎在子宫内的形态发育。我们制作了eG1-和G0-NT囊胚,然后将它们移植到受体小母牛体内,以便在子宫内短暂发育至妊娠第14天。体外受精(IVF)、孤雌生殖和人工授精(AI)胚胎用作对照。在eG1-NT、IVF和AI胚胎组中,回收胚胎的胚盘形成率相同(p>0.05)。eG1-NT胚胎的形成率显著高于G0-NT胚胎(p<0.05)。eG1-NT胚胎的长度与IVF、孤雌生殖和AI胚胎的长度相同(p>0.05),但显著短于G0-NT胚胎(p<0.01)。我们得出结论,源自eG1-NT胚胎的第14天胚胎的形态发育大多与AI胚胎相似,但G0-NT胚胎的形态发育异常大,且与AI和eG1-NT胚胎不同。