Jang Goo, Hong So Gun, Lee Byeong Chun
Department of Theriogenology and Biotechnology, College of Veterinary and the Research Institute of Veterinary Science, Seoul National University, Seoul 151-742, Korea.
J Vet Sci. 2011 Mar;12(1):83-9. doi: 10.4142/jvs.2011.12.1.83.
Somatic cell nuclear transfer (SCNT) is considered to be a critical tool for propagating valuable animals. To determine the productivity calves resulting from embryos derived with different culture media, enucleated oocytes matured in vitro were reconstructed with fetal fibroblasts, fused, and activated. The cloned embryos were cultured in modified synthetic oviduct fluid (mSOF) or a chemically defined medium (CDM) and developmental competence was monitored. After 7 days of culturing, the blastocysts were transferred into the uterine horn of estrus-synchronized recipients. SCNT embryos that were cultured in mSOF or CDM developed to the blastocysts stages at similar rates (26.6% vs. 22.5%, respectively). A total of 67 preimplantational stage embryos were transferred into 34 recipients and six cloned calves were born by caesarean section, or assisted or natural delivery. Survival of transferred blastocysts to live cloned calves in the mSOF and the CDM was 18.5% (to recipients), 9.6% (to blastocysts) and 42.9% (to recipients), 20.0% (to blastocysts), respectively. DNA analysis showed that all cloned calves were genetically identical to the donor cells. These results demonstrate that SCNT embryos cultured in CDM showed higher viability as judged by survival of the calves that came to term compared to blastocysts derived from mSOF cultures.
体细胞核移植（SCNT）被认为是繁殖珍贵动物的关键工具。为了确定用不同培养基培养的胚胎所产生的犊牛的生产率，将体外成熟去核的卵母细胞与胎儿成纤维细胞进行重构、融合并激活。将克隆胚胎在改良的合成输卵管液（mSOF）或化学成分明确的培养基（CDM）中培养，并监测其发育能力。培养7天后，将囊胚移植到发情同步受体的子宫角。在mSOF或CDM中培养的SCNT胚胎发育到囊胚阶段的比率相似（分别为26.6%和22.5%）。总共67个植入前阶段的胚胎被移植到34个受体中，通过剖腹产、辅助分娩或自然分娩出生了6头克隆犊牛。在mSOF和CDM中，移植的囊胚发育成活克隆犊牛的存活率分别为18.5%（相对于受体）、9.6%（相对于囊胚）和42.9%（相对于受体）、20.0%（相对于囊胚）。DNA分析表明，所有克隆犊牛在基因上与供体细胞相同。这些结果表明，与mSOF培养的囊胚相比，用CDM培养的SCNT胚胎根据足月出生犊牛的存活率判断具有更高的活力。
