广西巴马小香猪体细胞克隆

本研究旨在检验新生广西巴马小香猪肾脏成纤维细胞支持克隆胚胎完全的体内发育潜能,亦即能通过其构建出存活的克隆猪,从而为克隆技术在广西巴马小香猪资源保存和开发上的应用奠定基础。首先制备新生雄性广西巴马小香猪肾脏成纤维细胞,用其制备体细胞核移植胚胎,追踪观察体细胞核移植胚胎体外发育潜能,最后通过胚胎移植检验其完全的体内发育潜能。实验结果表明,制备的新生雄性广西巴马小香猪肾脏成纤维细胞具有良好的细胞增殖活性,用其制备的体细胞核移植胚胎分裂率和囊胚率分别为77.7%(334/430)和16.5%(71/430),将1 658枚克隆胚胎移植给6头代孕母猪,其中2头妊娠并最终产下8头存活雄性克隆小猪和3头死胎,整体克隆效率为0.66%,存活克隆猪健康状况良好。本研究表明,新生猪肾脏成纤维细胞是一种理想的用于生产体细胞克隆广西巴马小香猪的细胞资源。

Production of somatic cell cloned Guangxi Bama minipigs

This study is aimed to examine whether kidney fibroblast cells from newborn Guangxi Bama minipig can support completely in vivo development potential of somatic cell nuclear transfer (SCNT) derived embryos. The fibroblast cells were firstly isolated from one male newborn Guangxi Bama minipig, and then used as donor cells for producing SCNT embryos. The in vitro development of SCNT embryos was observed, and the SCNT embryos were transferred to surrogate sows for examining the completely in vivo development potential. In the present study, we established fibroblast cells from the kidney of one newborn Guangxi Bama minipig. The cells showed excellent proliferation activity. The fibroblast cells were then used as donor cells to produce SCNT embryos with the cleavage and blastocyst formation rates as 77.7%(334/430) and 16.5%(71/430), respectively. Finally, after transferred in total of 1 658 SCNT embryos to 6 surrogate recipients, we obtained 8 live cloned male piglets and 3 stillbirths. Overall cloning efficiency was 0.66%. All of the 8 piglets appeared healthy at birth and were observed to have normal growth after then. The present study indicates that the kidney fibroblast cells from newborn Guangxi Bama minipig can support the SCNT derived embryos achieving completely in vivo development potential, providing a ideal donor cell resource for producing cloned Guangxi Bama minipigs.