Development of Interspecies Cloned Monkey Embryos Reconstructed with Bovine Enucleated Oocytes
Development of Interspecies Cloned Monkey Embryos Reconstructed with Bovine Enucleated Oocytes
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DOI：
10.1262/jrd.20049
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发表时间：
2008-10-01
影响因子：
1.8
通讯作者：
Parnpai, Rangsun
中科院分区：
文献类型：
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作者：
Lorthongpanich, Chanchao;Laowtammathron, Chuti;Parnpai, Rangsun
本研究旨在确定用牛去核卵母细胞构建的培养液和Oct-4的表达模式是否支持猴成纤维细胞在异种克隆猴胚胎中的去分化。本研究以猴、牛皮肤成纤维细胞为供体细胞，进行牛去核卵母细胞移植重建。将重组猴种间体细胞核移植(ISCNT)胚胎在6种不同的培养条件下进行培养，改变胚胎培养液和正常牛、猴规格。用免疫细胞化学方法检测OCT-4在二细胞期至囊胚期的表达。猴iSCNT胚胎的卵裂率与牛SCNT和牛孤雌激活(PA)的卵裂率相似。然而，在任何培养条件下，猴子的iSCNT胚胎都不能发育到16-细胞期以上。在猴和牛的SCNT胚胎中，Oct-4在二细胞期到囊胚期都可以检测到，而在牛PA胚胎中，Oct-4从桑椹胚到囊胚期都可以检测到。这些结果表明，牛卵母细胞可以支持猴体细胞核的脱分化，但不能支持胚胎发育到致密桑拿期或囊胚期。综上所述，我们发现促进猴iSCNT胚胎发育的培养条件和Oct-4在克隆胚胎(猴iSCNT和牛SCNT)中的表达模式与在牛PA胚胎中的表达模式不同。
This study was carried out to determine whether Culture media reconstructed with bovine enucleated oocytes and the expression pattern of Oct-4 Could support dedifferentiaton of monkey fibroblasts in interspecies cloned monkey embryos. In this study, monkey and bovine skin fibroblasts were used as donor cells for reconstruction with bovine enucleated oocytes. The reconstructed monkey interspecies somatic cell nuclear transfer (iSCNT) embryos were then Cultured under six different Culture conditions with modifications of the embryo culture media and normal bovine and monkey specifications. The Oct-4 expression patterns of the embryos were examined at the two-cell to blastocyst stages using immunocytochemistry. The monkey iSCNT embroys showed similar cleavage rates to those of bovine SCNT and bovine parthenogenetic activation (PA). However, the monkey iSCNT embryos were not able to develop beyond the 16-cell stage under any of the culture conditions. In monkey and bovine SCNT embryos, Oct-4 could be detected front the two-cell to blastocyst stage, and in bovine PA embryos, Oct-4 was detectable from the morula to blastocyst stage. These results suggested that bovine ooplasm could Support dedifferentiation of monkey somatic Cell nuclei but could not support embryo development to either the compact morula or blastocyst stage. In conclusion, we found that the Culture conditions that tend to enhance monkey iSCNT embryo development and the expression pattern of Oct-4 in cloned embryos (monkey iSCNT and bovine SCNT) are different than in bovine PA embryos.