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利用家犬卵母细胞通过种间体细胞核移植成功克隆郊狼。

Successful cloning of coyotes through interspecies somatic cell nuclear transfer using domestic dog oocytes.

作者信息

Hwang Insung, Jeong Yeon Woo, Kim Joung Joo, Lee Hyo Jeong, Kang Mina, Park Kang Bae, Park Jung Hwan, Kim Yeun Wook, Kim Woo Tae, Shin Taeyoung, Hyun Sang Hwan, Jeung Eui-Bae, Hwang Woo Suk

机构信息

Sooam Biotech Research Foundation, 64 Kyungin-ro, Guro-gu, Seoul 152-895, Republic of Korea.

出版信息

Reprod Fertil Dev. 2013;25(8):1142-8. doi: 10.1071/RD12256.

Abstract

Interspecies somatic cell nuclear transfer (iSCNT) is an emerging assisted reproductive technology (ART) for preserving Nature's diversity. The scarcity of oocytes from some species makes utilisation of readily available oocytes inevitable. In the present study, we describe the successful cloning of coyotes (Canis latrans) through iSCNT using oocytes from domestic dogs (Canis lupus familiaris or dingo). Transfer of 320 interspecies-reconstructed embryos into 22 domestic dog recipients resulted in six pregnancies, from which eight viable offspring were delivered. Fusion rate and cloning efficiency during iSCNT cloning of coyotes were not significantly different from those observed during intraspecies cloning of domestic dogs. Using neonatal fibroblasts as donor cells significantly improved the cloning efficiency compared with cloning using adult fibroblast donor cells (P<0.05). The use of domestic dog oocytes in the cloning of coyotes in the present study holds promise for cloning other endangered species in the Canidae family using similar techniques. However, there are still limitations of the iSCNT technology, as demonstrated by births of morphologically abnormal coyotes and the clones' inheritance of maternal domestic dog mitochondrial DNA.

摘要

种间体细胞核移植(iSCNT)是一种新兴的辅助生殖技术(ART),用于保护自然界的生物多样性。某些物种的卵母细胞稀缺,使得利用容易获得的卵母细胞成为必然。在本研究中,我们描述了通过使用家犬(Canis lupus familiaris或澳洲野犬)的卵母细胞,经iSCNT成功克隆郊狼(Canis latrans)的过程。将320个种间重构胚胎移植到22只家犬受体体内,成功孕育了6只幼犬,最终产下8只存活后代。郊狼iSCNT克隆过程中的融合率和克隆效率与家犬种内克隆过程中观察到的情况无显著差异。与使用成年成纤维细胞供体细胞进行克隆相比,使用新生成纤维细胞作为供体细胞显著提高了克隆效率(P<0.05)。本研究中使用家犬卵母细胞克隆郊狼,为利用类似技术克隆犬科家族的其他濒危物种带来了希望。然而,iSCNT技术仍存在局限性,如形态异常的郊狼出生以及克隆体继承母本家犬线粒体DNA。

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