Chen Huanhuan, Zhang Lei, Guo Zekun, Wang Yongsheng, He Rongjun, Qin Yumin, Quan Fusheng, Zhang Yong
College of Veterinary Medicine, Northwest A & F University, Yangling, China.
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, China.
Mol Reprod Dev. 2015 Nov;82(11):867-79. doi: 10.1002/mrd.22531. Epub 2015 Aug 10.
Vitamin C (Vc) has been widely studied in cell and embryo culture, and has recently been demonstrated to promote cellular reprogramming. The objective of this study was to identify a suitable Vc concentration that, when used to treat adult bovine fibroblasts serving as donor cells for nuclear transfer, improved donor-cell physiology and the developmental potential of the cloned embryos that the donor nuclei were used to create. A Vc concentration of 0.15 mM promoted cell proliferation and increased donor-cell 5-hydroxy methyl cytosine levels 2.73-fold (P < 0.05). The blastocyst rate was also significantly improved after nuclear transfer (39.6% treated vs. 26.0% control, P < 0.05); the average number of apoptotic cells in cloned blastocysts was significantly reduced (2.2 vs. 4.4, P < 0.05); and the inner cell mass-to-trophectoderm ratio (38.25% vs. 30.75%, P < 0.05) and expression of SOX2 (3.71-fold, P < 0.05) and POU5F1 (3.15-fold, P < 0.05) were significantly increased. These results suggested that Vc promotes cell proliferation, decreases DNA methylation levels in donor cells, and improves the developmental competence of bovine somatic-cell nuclear transfer embryos.
维生素C（Vc）已在细胞和胚胎培养中得到广泛研究，最近被证明可促进细胞重编程。本研究的目的是确定一种合适的Vc浓度，当用于处理作为核移植供体细胞的成年牛成纤维细胞时，可改善供体细胞生理状态以及供体细胞核所构建的克隆胚胎的发育潜力。0.15 mM的Vc浓度可促进细胞增殖，并使供体细胞5-羟甲基胞嘧啶水平提高2.73倍（P < 0.05）。核移植后囊胚率也显著提高（处理组为39.6%，对照组为26.0%，P < 0.05）；克隆囊胚中凋亡细胞的平均数量显著减少（分别为2.2和4.4，P < 0.05）；内细胞团与滋养外胚层的比例（分别为38.25%和30.75%，P < 0.05）以及SOX2（3.71倍，P < 0.05）和POU5F1（3.15倍，P < 0.05）的表达显著增加。这些结果表明，Vc可促进细胞增殖，降低供体细胞中的DNA甲基化水平，并提高牛体细胞核移植胚胎的发育能力。
