Hwang Jeong Ho, Kim Sang Eun, Gupta Mukesh Kumar, Lee HoonTaek
1 Bio-Organ Research Center, Konkuk University , Seoul, Republic of Korea.
3 Jeonbuk Department of Inhalation Research, Korea Institute of Toxicology , Jeongeup, Republic of Korea.
Cell Reprogram. 2016 Aug;18(4):207-13. doi: 10.1089/cell.2015.0065.
Transgenic animal producing technology has improved consistently over the last couple of decades. Among the available methods, somatic cell nuclear transfer (SCNT) technology was officially the most popular. However, SCNT has low efficiency and requires a highly skilled individual. Additionally, the allo-SCNT nuclear reprogramming mechanism is poorly understood in the gnotobiotic miniature pig, which is a candidate for xenotransplantation, making sampling in oocytes very difficult compared to commercial hybrid pigs. Therefore, interbreed SCNT (ibSCNT), which is a combination of miniature pig and commercial pig (Landrace based), was analyzed and was found to be similar to SCNT in terms of the rate of blastocyst formation (12.6% ± 2.9% vs. 15.5% ± 2.2%; p > 0.05). However, a significantly lower fusion rate was observed in the ibSCNT compared to normal SCNT with Landrace pig somatic cells (29.6% ± 0.8% vs. 65.0% ± 4.9%). Thus, the optimization of fusion parameters was necessary for efficient SCNT. Our results further revealed that ibSCNT by the whole-cell intracytoplasmic injection (WCICI) method had a significantly higher blastocyst forming efficiency than the electrofusion method (31.1 ± 8.5 vs. 15.5% ± 2.2%). The nuclear remodeling and the pattern of changes in acetylation at H3K9 residue were similar in both SCNT and ibSCNT embryos.
在过去几十年中，转基因动物生产技术一直在不断改进。在现有的方法中，体细胞核移植（SCNT）技术是官方最常用的。然而，SCNT效率低下，需要高技能人员。此外，在无菌微型猪（异种移植的候选者）中，异体SCNT核重编程机制了解甚少，与商业杂交猪相比，在卵母细胞中取样非常困难。因此，对微型猪和商业猪（长白猪）杂交的体细胞核移植（ibSCNT）进行了分析，发现其囊胚形成率与SCNT相似（12.6%±2.9%对15.5%±2.2%；p>0.05）。然而，与使用长白猪体细胞的正常SCNT相比，ibSCNT的融合率明显较低（29.6%±0.8%对65.0%±4.9%）。因此，优化融合参数对于高效的SCNT是必要的。我们的结果进一步表明，通过全细胞胞质内注射（WCICI）方法的ibSCNT的囊胚形成效率明显高于电融合方法（31.1±8.5对15.5%±2.2%）。SCNT和ibSCNT胚胎中的核重塑以及H3K9残基的乙酰化变化模式相似。
