Cloned mice from fetal fibroblast cells arrested at metaphase by a serial nuclear transfer
Cloned mice from fetal fibroblast cells arrested at metaphase by a serial nuclear transfer
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DOI：
10.1095/biolreprod64.1.44
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发表时间：
2001-01-01
影响因子：
3.6
通讯作者：
Kono, T
中科院分区：
文献类型：
--
作者：
Ono, Y;Shimozawa, N;Kono, T
利用G（o）-阻滞的体细胞进行克隆，使人们认为细胞周期的这一阶段是克隆成功所必需的。本文报道了用细胞周期中期阻滞的胎儿成纤维细胞与去核的卵母细胞融合，孤雌激活后形成极体和单个二倍体原核，获得克隆小鼠。其中一些被允许发育到胚泡阶段，而另一些被去核，细胞核被转移到去核的受精1-细胞胚胎中。在单次移植技术后，164例妊娠晚期中有2例死亡，伴有大体异常。然而，在连续核移植后，272个胚胎中有5个在第19.5天存活，其中2个继续发育成明显正常的成年人。所有克隆胚胎均表现出严重的胎盘肥大和胎盘组织分化缺陷。这项研究表明，重编程可以发生在细胞周期中期的核移植后。
Cloning using G(o)-arrested somatic cells has led to the suggestion that this stage of the cell cycle is necessary for the success of cloning. In this study we report that cloned mice can be generated from fetal fibroblasts arrested at metaphase of the cell cycle, The procedure involves fusing a metaphase-arrested fetal fibroblast to an enucleated oocyte, After parthenogenetic activation a polar body and single diploid pronucleus were formed. Some of these were allowed to develop to the blastocyst stage, while others were enucleated and the nucleus was transferred to an enucleated fertilized 1-cell embryo. After the single transfer technique, 2 out of 164 developed to late stages of gestation were dead with gross abnormalities. However, after the serial nuclear transfer, 5 out of 272 embryos were recovered live at Day 19.5, and 2 of these went on to develop into apparently normal adults. All of the cloned embryos showed severe placental hypertrophy and defective differentiation of placental tissues. This study illustrates that reprogramming can occur after nuclear transfer at metaphase of the cell cycle.