Cloned ferrets produced by somatic cell nuclear transfer
Cloned ferrets produced by somatic cell nuclear transfer
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DOI：
10.1016/j.ydbio.2006.02.016
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发表时间：
2006-05-15
影响因子：
2.7
通讯作者：
Engelhardt, John F.
中科院分区：
文献类型：
--
作者：
Li, Ziyi;Sun, Xingshen;Engelhardt, John F.
体细胞核移植（SCNT）为开发更好的人类疾病动物模型提供了巨大的潜力。家养雪貂（Mustela putorius furo）是流感感染和其他潜在的人类呼吸系统疾病（如囊性纤维化）的理想动物模型，其中小鼠模型未能复制人类疾病表型。在这里，我们报告了使用物种特异性SCNT方法成功生产活克隆，生殖能力强的雪貂。为雪貂开发成功的SCNT方案的关键是发现通常用于超排卵的激素治疗会对受体卵母细胞的发育潜力产生不利影响。与与输精管切除的雄性自然交配的雌性相比，在激素处理的雌性孤雌激活的卵母细胞中，Oct4的表达延迟且不完全。交配和体外卵母细胞成熟诱导的刺激产生了SCNT的最佳卵母细胞受体。虽然核注射和细胞融合产生中期胎儿的比率相当（类似于3-4%），但只有细胞融合才能产生健康的存活克隆。将两个体细胞置于卵泡周围空间也能提高单细胞融合率和SCNT的效率。这些物种特有的修饰促进了活的、健康的、。还有可繁殖的克隆雪貂。对家养雪貂微卫星基因分型的研究证实，雪貂克隆是由它们各自的体细胞遗传而来，与它们的代孕母亲无关。有了这项技术，现在可以开始培育遗传定义的雪貂，用于研究传染性和遗传性人类肺部疾病。克隆家养雪貂也可能有助于恢复和保护濒危的黑足雪貂和欧洲水貂。(c) 2006爱思唯尔公司版权所有。
Somatic cell nuclear transfer (SCNT) offers great potential for developing better animal models of human disease. The domestic ferret (Mustela putorius furo) is an ideal animal model for influenza infections and potentially other human respiratory diseases such as cystic fibrosis, where mouse models have failed to reproduce the human disease phenotype. Here, we report the successful production of live cloned, reproductively competent, ferrets using species-specific SCNT methodologies. Critical to developing a successful SCNT protocol for the ferret was the finding that hormonal treatment, normally used for superovulation, adversely affected the developmental potential of recipient oocytes. The onset of Oct4 expression was delayed and incomplete in parthenogenetically activated oocytes collected from hormone-treated females relative to oocytes collected from females naturally mated with vasectomized males. Stimulation induced by mating and in vitro oocyte maturation produced the optimal oocyte recipient for SCNT. Although nuclear injection and cell fusion produced mid-term fetuses at equivalent rates (similar to 3-4%), only cell fusion gave rise to healthy surviving clones. Single cell fusion rates and the efficiency of SCNT were also enhanced by placing two somatic cells into the perivitelline space. These species-specific modifications facilitated the birth of live, healthy,. and fertile cloned ferrets. The development of microsatellite genotyping for domestic ferrets confirmed that ferret clones were genetically derived from their respective somatic cells and unrelated to their surrogate mother. With this technology, it is now feasible to begin generating genetically defined ferrets for studying transmissible and inherited human lung diseases. Cloning of the domestic ferret may also aid in recovery and conservation of the endangered black-footed ferret and European mink. (c) 2006 Elsevier Inc. All rights reserved.