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

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

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (31772554), the Department of Science and Technology of Guangdong Province, China (2016B020233006, 2015TX01N081 and 2016A020210074) and the team project of Guangdong Agricultural Department (2017LM2148).

Compliance with ethics guidelines

Chengcheng Zhao, Junsong Shi, Rong Zhou, Ranbiao Mai, Lvhua Luo, Xiaoyan He, Hongmei Ji, Gengyuan Cai, Dewu Liu, Enqin Zheng, Zhenfang Wu, and Zicong Li declare that they have no conflicts of interest or financial conflicts to disclose.

All applicable institutional and national guidelines for the care and use of animals were followed.

RIGHTS & PERMISSIONS

The Author(s) 2018. Published by Higher Education Press. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0)