Goto Yuji, Matoba Satoko, Imai Kei, Geshi Masaya
National Institute of Livestock and Grassland Science, Nagano, Japan.
J Reprod Dev. 2011 Apr;57(2):249-55. doi: 10.1262/jrd.10-102a. Epub 2010 Dec 22.
This study was conducted to improve the developmental ability of nuclear transfer (NT) embryos by using blastomeres from in vitro fertilized (IVF) embryos with high quality as donor cells. The IVF embryos selected at the 2-cell stage at 24-h postinsemination (hpi) and again at the ≥8-cell stage at 48 hpi (Selected-IVF-embryos) showed the highest blastocyst formation rate among embryos. When blastomeres from the Selected-IVF-embryos (Selected-NT group) or Nonselected-IVF-embryos (Non-selected-NT group) were used as donor cells for NT, the blastocyst formation rate in the Selected-NT group (25.6%) was significantly higher than that in the Non-selected-NT group (13.5%). When blastomeres from the Selected-IVF-embryos at 108 (contained many cells before cell division) and 126 hpi (contained many cells immediately after cell division) were used as donor cells for NT (108- and 126-NT groups, respectively), the 126-NT group showed a significantly higher blastocyst formation rate (32.1%) than the 108-NT group (16.8%). Embryo transfer of blastocysts in the 126-NT group showed that 11 of 23 recipients became pregnant; nine calves were obtained. For the NT embryos reconstructed using in vivo derived embryos, 9 of 20 recipients became pregnant; seven calves were obtained. These results indicate that the blastocyst formation rate of NT embryos can be improved by using blastomeres from IVF embryos selected at the early developmental stage, especially immediately after cell division, and that the resultant NT embryos have a high developmental ability to progress to term that is comparable to NT embryos reconstructed using in vivo derived embryos.
本研究旨在通过使用来自体外受精(IVF)优质胚胎的卵裂球作为供体细胞,提高核移植(NT)胚胎的发育能力。在授精后24小时(hpi)的2细胞阶段以及48 hpi时再次在≥8细胞阶段选择的IVF胚胎(选定的IVF胚胎)在胚胎中显示出最高的囊胚形成率。当来自选定的IVF胚胎(选定的NT组)或未选定的IVF胚胎(未选定的NT组)的卵裂球用作NT的供体细胞时,选定的NT组的囊胚形成率(25.6%)显著高于未选定的NT组(13.5%)。当来自108(细胞分裂前包含许多细胞)和126 hpi(细胞分裂后立即包含许多细胞)的选定IVF胚胎的卵裂球分别用作NT的供体细胞(分别为108-NT组和126-NT组)时,126-NT组的囊胚形成率(32.1%)显著高于108-NT组(16.8%)。126-NT组囊胚的胚胎移植显示,23只受体中有11只怀孕;获得了9头小牛。对于使用体内衍生胚胎重建的NT胚胎,20只受体中有9只怀孕;获得了7头小牛。这些结果表明,通过使用在早期发育阶段尤其是细胞分裂后立即选择的IVF胚胎的卵裂球,可以提高NT胚胎的囊胚形成率,并且由此产生的NT胚胎具有高发育能力,能够发育至足月,这与使用体内衍生胚胎重建的NT胚胎相当。