Hwang In-Sul, Kwon Dae-Jin, Oh Keun Bong, Ock Sun-A, Chung Hak-Jae, Cho In-Cheol, Lee Jeong-Woong, Im Gi-Sun, Hwang Seongsoo
Animal Biotechnology Division, National Institute of Animal Science, Wanju 565-851, Korea.
Subtropical Livestock Research Institute, NIAS, Jeju 690-150, Korea.
Dev Reprod. 2015 Jun;19(2):79-84. doi: 10.12717/DR.2015.19.2.079.
The Korean native pig (KNP) have been considered as animal models for animal biotechnology research because of their relatively small body size and their presumably highly inbred status due to the closed breeding program. However, little is reported about the use of KNP for animal biotechnology researches. This study was performed to establish the somatic cell nuclear transfer (SCNT) protocol for the production of swine leukocyte antigens (SLA) homotype-defined SCNT KNP. The ear fibroblast cells originated from KNP were cultured and used as donor cell. After thawing, the donor cells were cultured for 1 hour with 15 μM roscovitine prior to the nuclear transfer. The numbers of reconstructed and parthenogenetic embryos transferred were 98 ± 35.2 and 145 ± 11.2, respectively. The pregnancy and delivery rate were 3/5 (60%) and 2/5 (40%). One healthy SLA homotype-defined SCNT KNP was successfully generated. The recipient-based individual cloning efficiency ranged from 0.65 to 1.08%. Taken together, it can be postulated that the methodological establishment of the production of SLA homotype-defined cloned KNP can be applied to the generation of transgenic cloned KNP as model animals for human disease and xenotransplantation researches.
韩国本地猪(KNP)因其体型相对较小以及由于封闭育种计划可能具有高度近交状态,而被视为动物生物技术研究的动物模型。然而,关于将KNP用于动物生物技术研究的报道较少。本研究旨在建立用于生产猪白细胞抗原(SLA)同型定义的体细胞核移植(SCNT)KNP的体细胞克隆方案。源自KNP的耳成纤维细胞被培养并用作供体细胞。解冻后,供体细胞在核移植前用15μM的roscovitine培养1小时。移植的重构胚胎和孤雌胚胎数量分别为98±35.2和145±11.2。妊娠率和分娩率分别为3/5(60%)和2/5(40%)。成功产生了一头健康的SLA同型定义的SCNT KNP。基于受体的个体克隆效率在0.65%至1.08%之间。综上所述,可以推测,生产SLA同型定义的克隆KNP的方法学建立可应用于生成转基因克隆KNP,作为人类疾病和异种移植研究的模型动物。