Recloned dogs derived from adipose stem cells of a transgenic cloned beagle
Recloned dogs derived from adipose stem cells of a transgenic cloned beagle
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DOI：
10.1016/j.theriogenology.2010.11.035
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发表时间：
2011-04-15
期刊：
影响因子：
2.8
通讯作者：
Lee, Byeong Chun
中科院分区：
文献类型：
--
作者：
Oh, Hyun Ju;Park, Jung Eun;Lee, Byeong Chun
许多研究已经假设哺乳动物克隆的效率与供体细胞分化状态呈负相关，并且可以通过使用未分化的细胞作为核供体来提高。在这里，我们试图通过从转基因克隆的比格犬中核转移犬脂肪组织来源的间充质干细胞（cAd-MSCs）来重新克隆犬，以确定cAd-MSCs是否可以成为合适的供体细胞类型。为了分离cAd-MSC，从通过用红色荧光蛋白（RFP）基因遗传修饰的犬胎儿成纤维细胞的体细胞核移植（SCNT）产生的转基因克隆比格犬收集脂肪组织。cAd-MSCs表达RFP基因和MSCs的细胞表面标志物特征，包括CD 29、CD 44和thy1.1。此外，cAd-MSCs经历成骨，成脂，成肌，神经和软骨分化时，暴露于特定的分化诱导条件。为了研究cAd-MSCs的发育潜力，我们进行了SCNT。将融合对（82/109，75.2%）化学活化并转移到5只自然发情同步的替代品的输卵管中。其中一只（20%）保持妊娠，随后产下两只健康的克隆幼崽。本研究首次证明了通过cAd-MSCs的核转移成功生产克隆比格犬。本研究的另一个重要结果是通过来自转基因克隆比格犬的细胞的核转移成功地再克隆表达RFP的转基因克隆比格犬幼仔。总之，本研究表明，脂肪干细胞可以是一个很好的核供体来源的狗克隆。(C)2011 Elsevier Inc. All rights reserved.
A number of studies have postulated that efficiency in mammalian cloning is inversely correlated with donor cell differentiation status and may be increased by using undifferentiated cells as nuclear donors. Here, we attempted the recloning of dogs by nuclear transfer of canine adipose tissue-derived mesenchymal stem cells (cAd-MSCs) from a transgenic cloned beagle to determine if cAd-MSCs can be a suitable donor cell type. In order to isolate cAd-MSCs, adipose tissues were collected from a transgenic cloned beagle produced by somatic cell nuclear transfer (SCNT) of canine fetal fibroblasts modified genetically with a red fluorescent protein (RFP) gene. The cAd-MSCs expressed the RFP gene and cell-surface marker characteristics of MSCs including CD29, CD44 and thy1.1. Furthermore, cAd-MSCs underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. In order to investigate the developmental potential of cAd-MSCs, we carried out SCNT. Fused-couplets (82/109, 75.2%) were chemically activated and transferred into the uterine tube of five naturally estrus-synchronized surrogates. One of them (20%) maintained pregnancy and subsequently gave birth to two healthy cloned pups. The present study demonstrated for the first time the successful production of cloned beagles by nuclear transfer of cAd-MSCs. Another important outcome of the present study is the successful recloning of RFP-expressing transgenic cloned beagle pups by nuclear transfer of cells derived from a transgenic cloned beagle. In conclusion, the present study demonstrates that adipose stem cells can be a good nuclear donor source for dog cloning. (C) 2011 Elsevier Inc. All rights reserved.