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Abstract It has been widely suggested that the incompletely reprogramming of donor cell was the major reason of the inefficiency of nuclear transfer. The present study was to investigate the effect of histone deacetylase inhibitor(Scriptaid) on the reprogramming of yak(Bos grunniens) fetal fibroblast cells, and the in vitro developmental capability of yak-bovine(Bos grunniens) interspecies somatic cell nuclear transfer(iSCNT) embryos after donor cells were treated with Scriptaid. The yak fetal fibroblasts were treated with 0, 500, 1 000, 1 500 and 3 000 nmol/L Scriptaid for 24 h, respectively. Then observed cell morphology, accounted the survival rate, analyzed cell cycle by flow cytometry, detected acetylation level of H3K9ac by immunofluorescence, and evaluated the developmental competence of iSCNT embryos, which showed that the cell morphology was changed significantly, the cell became flatten and the area of nuclear was larger after Scriptaid treatment; cell proliferation was inhibited gradually and most cells were arrested at G0/G1 stage after Scriptaid treatment for 24 h; the acetylation level of H3K9ac was enhanced along with the concentrations of Scriptaid; the cleavage and blastocyst formation rate of iSCNT embryos improved significantly when donor cells treated with 1 000 nmol/L Scriptaid for 24 h(P<0.05). The results suggested that Scriptaid can significantly improve the histone acetylation level of yak fetal fibroblast cells, pre-treated donor cells with 1 000 nmol/L Scriptaid was helpful for improving in vitro development of yak-bovine iSCNT embryos. It was concluded that Scriptaid enhances the reprogramming in terms of high histone acetylation of donor cells, and it is useful for studying reprogramming and a novel way to improve the efficiency of yak iSCNT.
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Received: 05 June 2012
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