Effects of enucleation method on <i>in vitro</i> and <i>in vivo</i> development rate of cloned pig embryos

doi:10.15302/J-FASE-2018227

Front. Agr. Sci. Eng.

2019, Vol. 6

Issue (1) : 61-65 https://doi.org/10.15302/J-FASE-2018227

RESEARCH ARTICLE

Effects of enucleation method on in vitro and in vivo development rate of cloned pig embryos

Chengcheng ZHAO^1,², Junsong SHI³, Rong ZHOU³, Ranbiao MAI³, Lvhua LUO³, Xiaoyan HE³, Hongmei JI³, Gengyuan CAI^1,², Dewu LIU^1,², Enqin ZHENG^1,², Zhenfang WU^1,²(

), Zicong LI²(

)

¹. National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
². Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
³. Guangdong Wens Foodstuff Group Co., Ltd., Yunfu 527400, China

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Abstract

Enucleation is a crucial procedure for mammalian somatic cell nuclear transfer (SCNT), especially for domestic animal cloning. Oocytes of domestic animals such as pigs and cattle contain dark lipid droplets that hinder localization and removal of the nucleus. Using an oocyte enucleation technique that can obtain a high enucleation rate but has minimal negative effects on the reprogramming potential of oocyte for cloning is beneficial for enhancing the outcome of SCNT. In this study, we compared the pig cloning efficiency resulting from blind aspiration-based (BA-B) enucleation and spindle imaging system-assisted (SIS-A) enucleation, and compared the pig SCNT success rate associated with BA-B enucleation and blind aspiration plus post-enucleation staining-based (BAPPS-B) enucleation. SIS-A enucleation achieved a significantly higher oocyte enucleation success rate and tended to obtain a higher in vivo full term development rate of SCNT embryos than BA-B enucleation. BAPPS-B enucleation also obtained significantly higher in vitro as well as in vivo full term development efficiency of cloned porcine embryos than BA-B enucleation. These data indicate that SIS-A and BAPPS-B enucleation are better approaches for pig SCNT than BA-B enucleation.

Keywords cloning enucleation pig SCNT

Corresponding Author(s): Zhenfang WU,Zicong LI

Just Accepted Date: 10 May 2018 Online First Date: 04 June 2018 Issue Date: 25 February 2019

Cite this article:

Chengcheng ZHAO,Junsong SHI,Rong ZHOU, et al. Effects of enucleation method on in vitro and in vivo development rate of cloned pig embryos[J]. Front. Agr. Sci. Eng. , 2019, 6(1): 61-65.

URL:

https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2018227
https://academic.hep.com.cn/fase/EN/Y2019/V6/I1/61

Tab.1 Comparison of the success rates of BA-B and SIS-A enucleation

Tab.2 Comparison of in vitro development rate of pig SCNT embryos generated by BA-B and SIS-A enucleation

Tab.3 Comparison of in vivo development rate of pig SCNT embryos generated by the BA-B and SIS-A enucleation

Tab.4 Comparison of in vitro development rate of pig SCNT embryos generated by the BA-B and BAPPS-B enucleation

Tab.5 Comparison of in vivo development rate of pig SCNT embryos generated by the BA-B an BAPPS-B enucleation

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