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Journal of Animal and Veterinary Advances

Year: 2013  |  Volume: 12  |  Issue: 4  |  Page No.: 497 - 503

Study of Electro-Fusion/Activation in Somatic Cell Nuclear Transfer to Obtain Cloned Putian Black Pig Embryos

Hui Peng, Feng-Jun Liu, Xing-Feng Zhang, Yi-Fen Zhuang, Xiu-Ai Wang, Hong-Xiang Li, Zhi-Yong Hong, Xiu-Jiao Lin and Wen-Chang Zhang

Abstract

Putian Black pigs, an improved pig breed in China, are on the edge of extinction and urgently require protection. To establish a method of electro-fusion/activation for reconstructed embryos obtained by Somatic Cell Nuclear Transfer (SCNT), the applicability of microelectrode fusion in electro-fusion/activation of reconstructed embryos from SCNT and its optimal parameters were explored using pig oocyte maturation in vitro for 42-44 h as receptors and ear fibroblast as donors. Researchers also compared microelectrode fusion with traditional chamber fusion and the effects of different culture media (PZM-3 and NCSU-23) on reconstructed embryos were evaluated. Researchers found that the optimal parameters for microelectrode fusion/activation were 16 V, 20 μsec and 1 DC in which the fusion efficiency was significantly higher than that of chamber-fusion (84.8 vs. 64.4%) but the rates of embryo cleavage and blastocyst formation showed no statistical difference (p>0.05). The cleavage rates of embryos cultured in PZM-3 or NCSU-23 showed no statistical difference (p>0.05) while the rate of blastocyst formation in PZM-3 was higher than that in NCSU-23 (13.8 vs. 7.84%, p<0.05). These results show that microelectrode fusion/activation can be used in SCNT to obtain cloned embryos from Putian Black pigs and its fusion efficiency is higher than that of traditional chamber fusion. In addition, reconstructed embryo development is optimal in PZM-3. This study establishes a technical basis for SCNT to obtain cloned embryos from Putian Black pigs. Moreover, it can be applied to related fields such as the production of transgenic cloned embryos from Putian Black pigs.

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