全细胞胞质内注射法生产猪重构胚的研究(简报)  

PRODUCTION OF PORCINE RECONSTRUCTED EMBRYOS BY WHOLE-CELL INTRACYTOPLASMIC MICROINJECTION

作　　者：孔庆然[1] 罗一博[1] 田江天[1] 王振坤[1] 张力[1] 刘忠华[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《分子细胞生物学报》2008年第1期70-74,共5页Journal of Molecular Cell Biology

摘　　要：自从1997年克隆羊“Dolly”出生以来。体细胞核移植技术得到广泛应用,已经有十余种克隆哺乳动物出生，但克隆效率仍然很低。核移植方法的不够完善是其重要原因之一。传统的核移植方法——电融合法和胞质内注射法虽然都取得了成功。但还存在操作复杂及需要辅助设备等问题。为了能够找出一种既容易操作，又不需要特殊设备的核移植方法，本研究对胞质内注射技术进行了改进,尝试了一种新的核移植方法——非Piezo辅助的全细胞胞质内注射法，即不通过Piezo辅助。不破坏供体细胞膜直接将供体细胞注入卵母细胞内，并且比较了这种方法与电融合法对猪体细胞核移植效率的影响。探讨了非Piezo辅助的全细胞胞质内注射法在猪体细胞核移植上的可行性。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.

关 键 词：猪 体细胞核移植 电融合法 全细胞胞质内注射法 

分 类 号：Q813[生物学—生物工程]