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Frontiers of Agricultural Science and Engineering

ISSN 2095-7505

ISSN 2095-977X(Online)

CN 10-1204/S

邮发代号 80-906

Frontiers of Agricultural Science and Engineering  2019, Vol. 6 Issue (1): 61-65   https://doi.org/10.15302/J-FASE-2018227
  本期目录
Effects of enucleation method on in vitro and in vivo development rate of cloned pig embryos
Chengcheng ZHAO1,2, Junsong SHI3, Rong ZHOU3, Ranbiao MAI3, Lvhua LUO3, Xiaoyan HE3, Hongmei JI3, Gengyuan CAI1,2, Dewu LIU1,2, Enqin ZHENG1,2, Zhenfang WU1,2(), 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.

Key wordscloning    enucleation    pig    SCNT
收稿日期: 2017-12-07      出版日期: 2019-02-25
Corresponding Author(s): Zhenfang WU,Zicong LI   
 引用本文:   
. [J]. Frontiers of Agricultural Science and Engineering, 2019, 6(1): 61-65.
Chengcheng ZHAO, Junsong SHI, Rong ZHOU, Ranbiao MAI, Lvhua LUO, Xiaoyan HE, Hongmei JI, Gengyuan CAI, Dewu LIU, Enqin ZHENG, Zhenfang WU, Zicong LI. Effects of enucleation method on in vitro and in vivo development rate of cloned pig embryos. Front. Agr. Sci. Eng. , 2019, 6(1): 61-65.
 链接本文:  
https://academic.hep.com.cn/fase/CN/10.15302/J-FASE-2018227
https://academic.hep.com.cn/fase/CN/Y2019/V6/I1/61
Enucleation method Number of manipulated/successfully enucleated oocytes Enucleation rate/%
BA-B 384/319 83.07**
SIS-A 322/306 95.03**
Tab.1  
Enucleation method SCNT embryos Cleaved/% Blastocysts/% Total number of cells per blastocyst
BA-B 240 183 (76.25) 44 (18.33) 40±3.34
SIS-A 279 208 (74.55) 37 (13.26) 47±4.10
Tab.2  
Enucleation method Total number of transferred SCNT embryos Total number/pregnant/farrowed recipients Pregnancy/farrowing rate/% Total number of cloned piglets born Development rate/%
BA-B 7469 33/23/14 69.70/42.42 64 0.86
SIS-A 6779 33/23/14 69.70/42.42 75 1.11*
Tab.3  
Enucleation method SCNT embryos Cleaved/% Blastocysts/% Total number of cells per blastocyst
BA-B 429 326 (75.99) 94 (21.91) 36±1.77*
BAPPS-B 501 362 (72.26) 89 (17.76) 41±1.69*
Tab.4  
Enucleation method Number of manipulated /successfully enucleated oocytes Total number of transferred SCNT embryos Total number/pregnant/farrowed recipients Pregnancy/farrowing rate/% Total number of born cloned piglets Development rate/%
BA-B 8023/not determined 7038 31/18/7 58.06/22.58* 24 0.34**
BAPPS-B 9049/7234 6735 31/21/15 67.74/48.39* 55 0.82**
Tab.5  
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