Production of transgenic and non-transgenic clones in miniature pigs by somatic cell nuclear transfer
Production of transgenic and non-transgenic clones in miniature pigs by somatic cell nuclear transfer
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DOI：
10.1262/jrd.19165
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发表时间：
2008-06-01
影响因子：
1.8
通讯作者：
Nagashima, Hiroshi
中科院分区：
文献类型：
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作者：
Kurome, Mayuko;Ishikawa, Takayuki;Nagashima, Hiroshi
小型猪在医学和药学研究等各个领域都被认为是有价值的实验动物。然而，关于小型猪和转基因小型猪体细胞克隆的信息量远远少于普通家猪。本研究旨在建立一种高效的体细胞核移植技术来克隆小型猪。以妊娠早期雌性小型猪为受体，用前列腺素F2α类似物和促性腺激素同步发情，移植孤雌胚胎(3/3)和克隆胚胎(5/6)，获得了较高的妊娠率。雄性和雌性胎儿成纤维细胞作为供核细胞克隆小型猪的生产效率分别为0.9%(2/215)和3/331(P<0.05)。将重构胚胎移植到普通家猪的子宫内，克隆小型猪的成活率也达到了7.8%(5/)。将转绿色荧光蛋白(GFP)基因的供体细胞用于核移植，重构胚胎的生产效率和囊胚发育率与未转染细胞相当。将转基因细胞来源的重组胚胎移植给3头普通家猪受体，全部受孕，共获得10头转基因克隆小型猪(仔猪生产效率：2.7%，10/365)。因此，我们能够建立一套实用的克隆和转基因克隆同基因背景小型猪的体系。
Miniature pigs have been recognized as valuable experimental animals in various fields such as medical and pharmaceutical research. However, the amount of information on somatic cell cloning in miniature pigs, as well as genetically modified miniature pigs, is much less than that available for common domestic pigs. The objective of the present study was to establish an efficient technique of cloning miniature pigs by somatic cell nuclear transfer. A high pregnancy rate was achieved following transfer of parthenogenetic (3/3) and cloned (5/6) embryos using female miniature pigs in the early pregnancy period as recipients after estrus synchronization with prostaglandin F2 alpha analog and gonadotrophins. The production efficiency of the cloned miniature pigs using male and female fetal fibroblasts as nucleus donors was 0.9% (2/215 and 3/331, respectively). Cloned miniature pigs were also produced efficiently (7.8%, 5/64) by transferring reconstructed embryos into the uteri of common domestic pigs. When donor cells transfected with the green fluorescent protein (GFP) gene were used in nuclear transfer, the production efficiency of the reconstructed embryos and rate of blastocyst development were comparable to those obtained by non-transfected cells. When transfected cell-derived reconstructed embryos were transferred to three common domestic pig recipients, all became pregnant, and a total of ten transgenic cloned miniature pigs were obtained (piglet production efficiency: 2.7%, 10/365). Hence, we were able to establish a practical system for producing cloned and transgenic-cloned miniature pigs with a syngeneic background.