5-Aza-CdR对德保黑猪手工克隆重构胚胎体外发育效果的影响

doi:10.16431/j.cnki.1671-7236.2018.11.020

Abstract

Abstract:

To investigate the effect of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on in vitro developmental potency of handmade clone (HMC) reconstructed embryos in Debao Black pigs,this study was performed from donor cells and reconstructed embryos,compared with developmental effects of 5-Aza-CdR treatment with HMC reconstructed embryos of five different concentrations (0,5,10,20 and 40 nmol/L) and five different treatment times (0,24,48,72,and 96 h),and screened for the optimal treatment concentration and time;Compared with in vitro developmental potency of four different concentrations (0,0.25,0.5 and 1 μmol/L) 5-Aza-CdR,combined with optimal concentration and optimal time to treatment with donor cells and reconstructed embryos.The results showed that there was no significant difference in the cleavage rate of reconstructed embryos between 5,10,20 and 40 nmol/L 5-Aza-CdR for 72 h compared with blank control group (P>0.05),20 nmol/L 5-Aza-CdR group significantly increased the blastocyst rate of reconstructed embryos (P<0.05).Both 10 and 20 nmol/L 5-Aza-CdR significantly increased the number of blastocysts (P<0.05),20 nmol/L 5-Aza-CdR was the best.Compared with blank control group,treatment with 20 nmol/L 5-Aza-CdR for 72 h significantly increased the blastocyst rate and blastocyst number of the reconstructed embryos (P<0.05),the remaining treatment time had no significant effect on the cleavage rate,blastocyst rate and blastocyst number of reconstructed embryos (P>0.05).Under optimal treatment concentration (20 nmol/L) and optimal treatment time(72 h),combined with four treatment concentrations (0,0.25,0.5 and 1 μmol/L) of the donor cells and treated with the reconstructed embryos and the donor cells simultaneously,the developmental potency of HMC reconstructed embryos in each treatment group was promoted,but the effect was not significant (P>0.05),which 0.25 to 0.5 μmol/L 5-Aza-CdR was better.In summary,the appropriate concentration (0.25 to 0.5 μmol/L) treatment of donor cells with DNA methylase inhibitor 5-Aza-CdR and treatment of reconstructed embryos with 20 nmol/L 5-Aza-CdR for 72 h could effectively improve in vitro developmental potency of HMC reconstructed embryos in Debao Black pig,which laid a foundation for the future study of DNA methylation regulation mechanism of HMC embryo in Debao Black pig.

Key words: 5-Aza-2'-deoxycytidine (5-Aza-CdR); Debao Black pig; handmade clone (HMC); developmental potency

CLC Number:

S814.8

LYU Lingyan, LU Xingrong, SUN Junming, CHEN Baojian, WU Yongshao, SUN Ruyu, WANG Yanling, CUI Kuiqing. Effect of 5-Aza-CdR on the in vitro Developmental Potency of Reconstructed Embryos of Debao Black Pig Handmade Clone[J]. , 2018, 45(11): 3144-3152.

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Effect of 5-Aza-CdR on the in vitro Developmental Potency of Reconstructed Embryos of Debao Black Pig Handmade Clone

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