Mice cloned from embryonic stem cells
Mice cloned from embryonic stem cells
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DOI：
10.1073/pnas.96.26.14984
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发表时间：
1999-12-21
影响因子：
11.1
通讯作者：
Mombaerts, P
中科院分区：
文献类型：
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作者：
Wakayama, T;Rodriguez, I;Mombaerts, P
克隆允许选定的个体进行无性繁殖，从而使后代拥有基本上相同的核基因组。到目前为止，通过核移植进行克隆的报道仅限于新鲜分离的细胞和来自原代培养的细胞。我们之前报道了一种通过显微注射从成体细胞核移植后克隆小鼠的方法。在这里，我们应用这种方法从广泛可用的已建立的胚胎干细胞系中克隆小鼠，在后期传代。在ES细胞系R1中，29%的重构卵母细胞在体外发育到桑椹/囊胚期，其中8%的胚胎移植给代孕母亲后发育成活体幼崽。因此，我们从R1细胞中克隆了26只小鼠。来自ES细胞系E14的细胞核也被显示出直接发育到足月。我们提出的证据表明，处于G(1)期或C-2/M期的ES细胞的核能够有效地支持完全发育。我们的发现表明，后期传代的ES细胞可以用来制造可存活的克隆小鼠，并提供了ES细胞技术和动物克隆之间的联系。因此，在较长时期内从单个细胞克隆大量个体是可能的。
Cloning allows the asexual reproduction of selected individuals such that the offspring have an essentially identical nuclear genome. Cloning by nuclear transfer thus far has been reported only with freshly isolated cells and cells from primary cultures. We previously reported a method of cloning mice from adult somatic cells after nuclear transfer by microinjection. Here, we apply this method to clone mice from widely available, established embryonic stem (ES) cell lines at late passage. With the ES cell line R1, 29% of reconstructed oocytes developed in vitro to the morula/blastocyst stage, and 8% of these embryos developed to live-born pups when transferred to surrogate mothers. We thus cloned 26 mice from R1 cells. Nuclei from the ES cell line E14 also were shown to direct development to term. We present evidence that the nuclei of ES cells at G(1)- or C-2/M-phases are efficiently able to support full development. Our findings demonstrate that late-passage ES cells can be used to produce viable cloned mice and provide a link between the technologies of ES cells and animal cloning. It thus may be possible to clone from a single cell a large number of individuals over an extended period.