SummaryThis study was performed to improve production efficiency at the level of recipient pig and donor nuclei of transgenic cloned pigs used for xenotransplantation. To generate transgenic pigs, human endothelial protein C receptor (hEPCR) and human thrombomodulin (hTM) genes were introduced using the F2A expression vector into GalT–/–/hCD55+ porcine neonatal ear fibroblasts used as donor cells and cloned embryos were transferred to the sows and gilts. Cloned fetal kidney cells were also used as donor cells for recloning to increase production efficiency. Pregnancy and parturition rates after embryo transfer and preimplantation developmental competence were compared between cloned embryos derived from adult and fetal cells. Significantly higher parturition rates were shown in the group of sows (50.0 vs. 4.1%), natural oestrus (20.8 vs. 0%), and ovulated ovary (16.7 vs. 5.6%) compared with gilt, induced and non-ovulated, respectively (P < 0.05). When using gilts as recipients, final parturitions occurred in only the fetal cell groups and significantly higher blastocyst rates (15.1% vs. 21.3%) were seen (P < 0.05). Additionally, gene expression levels related to pluripotency were significantly higher in the fetal cell group (P < 0.05). In conclusion, sows can be recommended as recipients due to their higher efficiency in the generation of transgenic cloned pigs and cloned fetal cells also can be recommended as donor cells through correct nuclear reprogramming.

中文翻译：

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通过重新克隆和使用母猪作为受体来提高多基因修饰猪的生产效率

总结本研究旨在提高用于异种移植的转基因克隆猪的受体猪和供体细胞核水平的生产效率。为了产生转基因猪，人内皮蛋白 C 受体 (hEPCR) 和人血栓调节蛋白 (hTM) 使用 F2A 表达载体将基因导入高尔特–/–/hCD55+将用作供体细胞的猪新生耳成纤维细胞和克隆胚胎移植到母猪和后备母猪身上。克隆的胎儿肾细胞也被用作重新克隆的供体细胞以提高生产效率。比较了来自成人和胎儿细胞的克隆胚胎之间的胚胎移植后的怀孕和分娩率以及植入前的发育能力。与后备母猪、诱导和未排卵母猪相比，母猪组（50.0 对 4.1%）、自然发情组（20.8 对 0%）和排卵卵巢组（16.7 对 5.6%）的分娩率分别显着更高(磷< 0.05)。当使用后备母猪作为受体时，最终分娩仅发生在胎儿细胞组中，并且胚泡率显着提高（15.1% 对 21.3%）。磷< 0.05)。此外，与多能性相关的基因表达水平在胎儿细胞组中显着更高。磷< 0.05)。总之，可以推荐母猪作为受体，因为它们在产生转基因克隆猪和克隆胎儿细胞方面的效率更高，也可以通过正确的核重编程推荐作为供体细胞。