Application of the modified handmade cloning technique to pigs

Eun Ji Lee1, Kuk Bin Ji1, Hyun Ju Oh2, Ji Hye Lee1, Tae Young Kil3, Min Kyu Kim1,2,*
Author Information & Copyright
1Division of Animal and Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon 34134, Korea.
2MK biotech Inc., 99 Daehak-ro, Yuseong-gu, Daejeon 34134, Korea.
3Department of Social Welfare, Joongbu University, 201 Daehak-ro, Geumsan-gun, Chungnam 32713, Korea.
*Corresponding Author: Min Kyu Kim, Division of Animal and Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon 34134, Korea, Republic of. MK biotech Inc., 99 Daehak-ro, Yuseong-gu, Daejeon 34134, Korea, Republic of. E-mail:

© Copyright 2021 Korean Society of Animal Science and Technology. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: Dec 21, 2020; Revised: Jan 28, 2021; Accepted: Jan 28, 2021

Published Online: Feb 05, 2021


Although somatic cell nuclear transfer (SCNT) is frequently employed to produce cloned animals in laboratories, this technique is expensive and inefficient. Therefore, the handmade cloning (HMC) technique has been suggested to simplify and advance the cloning process, however, HMC wastes many oocytes and leads to mitochondrial heteroplasmy. To solve these problems, we propose a modified handmade cloning (mHMC) technique that uses simple laboratory equipment, i.e., a Pasteur pipette and an alcohol lamp, and apply it to porcine embryo cloning. To validate the application of mHMC to pig cloning, embryos produced through SCNT and mHMC are compared using multiple methods, such as enucleation efficiency, oxidative stress, embryo developmental competence, and gene expression. The results show no significant differences between techniques except in the enucleation efficiency. The 8-cell and 16-cell embryo developmental competence and Oct4 expression levels exhibit significant differences. however, the blastocyst rate is not significantly different between mHMC and SCNT. This study verifies that cloned embryos derived from the two techniques exhibit similar generation and developmental competence. thus, we suggest that mHMC could replace SCNT for simpler and cheaper porcine cloning.

Keywords: somatic cell nuclear transfer; in vitro culture; modified handmade cloning; porcine embryo

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