全细胞胞浆内显微注射制备猪重构胚胎。

Qing Ran Kong, Yi Bo Luo, Jiang Tian Tian, Zhen Kun Wang, Li Zhang, Zhong Hua Liu
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引用次数: 0

摘要

体细胞核移植克隆已通过电融合和胞浆内核注射(ICNI)两种方法实现。然而,上述每种方法都涉及到扩展的复杂操作和特殊设备。在这里,我们报告了一种没有压电辅助的全细胞注射技术用于猪的核移植。以猪成纤维细胞为核供细胞,研究了新方法对猪体细胞核移植效率的影响,并与电融合法进行了比较。结果表明,两种方法的重构胚制备效率略低,但差异不显著(88.4% vs 78%, P > 0.05)。胚胎培养48h和7d后,融合法在卵母细胞分裂率和囊胚发育方面均优于新方法(78% vs 53.2%, P < 0.05;27.2% vs 13.8%, P < 0.01)。结果表明,两种方法对囊胚的质量没有影响,但电融合方法效率更高。因此,通过简单且劳动强度低的全细胞胞浆内注射生产正常克隆仔猪的适用性有待进一步提高。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Production of porcine reconstructed embryos by whole-cell intracytoplasmic microinjection].

Cloning by somatic cell nuclear transfer has been achieved by both electric fusion and intracytoplasmic nuclear injection (ICNI) methods. However, each of the above methods involves extended complicate manipulation and special equipment. Here we report a whole-cell injection technique without Piezo assistance for nuclear transfer in pigs. The fibroblast cell of pig as the nucleus donor cell, effects of the new method on the efficiency of somatic nuclear transfer in pig were investigated, compared with that of electric fusion method. Results showed that the new method was a little less efficient in producing reconstructed embryos but without significant difference (88.4% vs 78%, P > 0.05). After the embryos were cultured 48h and 7d, the fusion method is more efficient than the new method in the oocyte cleavage rate and the blastocyst development (78% vs 53.2%, P < 0.05; 27.2% vs 13.8%, P < 0.01). The results indicate that both methods make no difference in the quality of the blastula, but the electric fusion method is more efficient. Therefore, the applicability of producing normal,cloned piglets by the simple and less labor-intensive whole-cell intracytoplasmic injection needs further improvement.

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