Effect Of Scriptaid On Embryonic Development Of Somatic Cell Nuclear Transfer Swine

Somatic cell nuclear transplantation(SCNT)is a powerful technique which is benefit for animal reproduction and breeding,local germplasm resources preservation,extincted animal preservation and it also plays an important role for the solution of many basic biological problems.However,the overall efficiency of SCNT is very low,Pregnancy abortion rate is high,donor and receptor inconsistent synchronous coordination,cloned animals phenotypic normal and abnormal essence,theory and technology is not mature,this problem has become the main hurdle that restricted the application of the technique.Therefore,it is necessary for us to optimize the protocal of SNT technique and investigate its mechanisms fully and completely.This study is dedicated to the detection of a new low protein group of different concentrations and time of treatment on the cloning of porcine embryos deacetylase inhibitor Scriptaid,and then the cleavage rate and in vitro embryo development ability.On the basis of the control experiment,the porcine fetal fibroblasts in Lingyuan Hefeng pig farm were used as donor cells,and then the embryos were obtained by somatic cell nuclear transfer(SCNT).The reconstructed embryos with different concentrations(0,200,500,700 and 900 nmol/L)after Scriptaid treatment,and different time(0,15,36 and 72 h)training,and then through the observation of different concentrations and treatment time in the 2 cell stage embryo cleavage rate and the blastocyst rate.The relative expression of Oct4 and Sox2 two factors in the control group and Scriptaid treated group were detected by Real-time PCR in the 2 cell stage.Through statistical analysis,the results showed that compared with the non treatment group,500 nmol/L Scriptaid treatment of blastocyst formation rate increased significantly(Vs.15.5% = 27%)(P < 0.05).2 cell division rate unchanged,while the untreated group of cloned embryos in multiple blastocyst period Oct4 gene expression was significantly lower than that of cloned embryos at the blastocyst stage after Scriptaid treatment the expression ratio of P < 0.05,indicating that the 500nmol/L after Scriptaid treatment,the cloned embryos Oct4 expression increased significantly.Without the expression of cloned embryos at the blastocyst stage with multiple Sox2 gene and cloned embryos treated with Scriptaid expression after multiple Sox2 gene in blastocyst stage are different but the difference is not obvious,this shows that after 500nmol/L Scriptaid treatment,the expression of Sox2 gene although there are improved,but overall and untreated embryos showed little difference P > 0.05.