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Frontiers of Agricultural Science and Engineering

ISSN 2095-7505

ISSN 2095-977X(Online)

CN 10-1204/S

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Front. Agr. Sci. Eng.    2019, Vol. 6 Issue (1) : 73-80    https://doi.org/10.15302/J-FASE-2018220
RESEARCH ARTICLE
Factors influencing the somatic cell nuclear transfer efficiency in pigs
Yong JIN, Manling ZHANG, Xinrong JU, Shuang LIANG, Qiang XIONG, Lihua ZHAO, Xiaowei NIE, Daorong HOU, Qiang LIU, Junzheng WANG, Chenyu WANG, Xiaokang LI, Lining ZHANG, Xiaorui LIU, Ying WANG, Haiyuan YANG, Yifan DAI(), Rongfeng LI()
Jiangsu Key Laboratory of Xenotransplantation/Key Laboratory of Targeted Intervention of Cardiovascular Disease/Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing 211166, China
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Abstract

Using a data set from our laboratory, we assessed the effects of several factors on pig cloning efficiency. The results demonstrated that cells at high confluence (>90%) used as donor cell resulted in higher pregnancy rate, delivery rate and overall cloning efficiency (number of live offspring born per reconstructed embryo transferred to recipients) compared with the cells at 60% to 79% confluence and 80% to 89% confluence. Cells with four, five and six passages compromised the pregnancy and delivery rates compared with first passage cells. The number of blastocysts transferred by somatic cell nuclear transfer (SCNT) did not significantly affect the cloning efficiency, but transfer of blastocyst derived from in vitro culture 5 d after SCNT achieved a significantly higher pregnancy rate compared with one to two cell SCNT embryos from overnight culture. The highest pregnancy rate, delivery rate and the largest litter size were obtained when Bama Miniature pig fibroblasts were used as donor cells and Landrace/Yorkshire hybrid gilts were used as recipients. Recipients treated with chemicals for estrus synchronization had higher pregnancy rates compared with untreated recipients. Our data might be helpful for improving SCNT efficiency in pigs.
Keywords blastocyst      donor cell      estrus synchronization      pregnancy rate      pig cloning      somatic cell nuclear transfer     
Corresponding Author(s): Yifan DAI,Rongfeng LI   
Just Accepted Date: 10 April 2018   Online First Date: 10 May 2018    Issue Date: 25 February 2019
 Cite this article:   
Yong JIN,Manling ZHANG,Xinrong JU, et al. Factors influencing the somatic cell nuclear transfer efficiency in pigs[J]. Front. Agr. Sci. Eng. , 2019, 6(1): 73-80.
 URL:  
https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2018220
https://academic.hep.com.cn/fase/EN/Y2019/V6/I1/73
Cell culture time(conflue-nce degree) No. of transferred embryos No. of recipients No. of pregnant recipients/% No. of farrowed recipients/% No. of piglets/% Litter size
0 d 340 7 3 (42.9)a 3 (42.9)a 13 (3.8)a 4.3
1–2 d (60%–79%) 933 16 1 (6.3)b 1 (6.3)b 5 (0.5)b 5.0
3 d (80%–89%) 757 12 1 (8.3)b 1 (8.3)b 5 (0.7)b 5.0
4 d (90%–100%) 582 9 4 (44.4)a 2 (22.2)a b 12 (2.1)a b 6.0
Tab.1  The effect of the culture time and confluence degrees of donor cells on pig cloning efficiency
Cell passage No. of transferred embryos No. of recipients No. of pregnant recipients/% No. of farrowed recipients/% No. of piglets/% Litter size
1 561 6 4 (66.7)a 4 (66.7)a 14 (2.5)a 3.5
4 773 11 2 (40.0)b 2 (10.0)c 8 (0.4)b 3.0
5 486 8 3 (42.9) b 3 (42.9)b 16 (4.1)a 5.7
6 546 9 3 (44.4)b 3 (44.4)b 13 (2.6)a 3.5
7 286 5 0 0 0 0
8 292 4 0 0 0 0
Tab.2  The effect of cell passage on transgenetic pigs production
Fig.1  The ovulation statuses of recipient. The SCNT embryos cultured for 22–24 h (a), 44–48 h (b) and 5 d (c) were transferred to recipients with different ovulation statuses, respectively.
Embryo culture time No. of transferred embryos No. of recipients No. of pregnant recipients/% No. of farrowed recipients/% Mean litter size
22–24 h 2993 10 2 (20.0)a 1 (10.0) 11.0
44–48 h 3719 12 6 (50.0)ab 2 (16.7) 6.5
5 d 704 11 9 (81.8)b 4 (36.4) 2.8
Tab.3  The effect of cloned embryo culture time on pig cloning efficiency
Transferred embryos per recipient No. of transferred embryos No. of recipients No. of pregnant recipients/% No. of farrowed recipients/% No. of piglets/% Litter size
30–49 421 12 4 (33.3) 2 (16.7) 8 (1.9) 4.0
50–69 1608 27 11 (40.7) 5 (18.5) 21 (1.3) 4.2
70–110 1745 23 8 (34.8) 5 (21.7) 19 (2.1) 3.8
Tab.4  The effect of transferred blastocyst number on pig cloning efficiency
Donor fibroblast cell type Recipient’s breeds No. of transferred embryos No. of recipients No. of pregnant recipients/% No. of farrowed recipients/% No. of piglets/% Mean litter size
LY-FBs Meishan 2289 16 2 (12.5)a 2 (12.5)a 8 (0.4) 4
LY-FBs LY 3747 18 4 (22.2)a 4 (22.2)a b 14 (0.4) 3.5
BM-FBs LY 3837 13 7 (53.9)b 6 (46.2)b 36 (0.9) 6
Tab.5  The effect of donor fibroblast cell and recipient’s breeds on pig cloning efficiency
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