16 HISTONE DEACETYLASE INHIBITORS PCI-24781 AND QUISINOSTAT IMPROVE THE IN VITRO DEVELOPMENTAL COMPETENCE OF PIG SOMATIC CELL NUCLEAR TRANSFER EMBRYOS
L. Jin A , H.-Y. Zhu A , Q. Guo A , Y.-C. Zhang A , X.-C. Li A , J.-D. Kang A and X.-J. Yin AYanbian University Jilin Provincial Key Laboratory of Transgenic Animal and Embryo Engineering, Yanji, Jilin, China
Reproduction, Fertility and Development 28(2) 138-138 https://doi.org/10.1071/RDv28n2Ab16
Published: 3 December 2015
Abstract
The aim of this study was to examine the effects of PCI-24781 and quisinostat, two novel histone deacetylase inhibitors, on the in vitro development of pig somatic cell NT (SCNT) embryos. Pig fetal fibroblasts were used as donor cells for SCNT embryos. In vitro-matured eggs with first polar body were enucleated by aspiration using a 15-μm inner diameter glass pipette. A single donor cell was inserted into the perivitelline space of each egg and electrically fused using 2 direct pulses of 150 V mm–1 for 50 μs in 0.28 M mannitol. After 1 h, embryos were activated by 2 direct pulses of 100 V mm–1 for 20 μs and incubated with 2 mM 6-DMAP for 4 h. Subsequently, the cloned embryos were cultured in medium for 7 days. In Experiment 1, after activation and treatment with 6-DMAP for 4 h, pig SCNT embryos were treated with various concentrations of PCI-24781 or quisinostat for 24 h. In Experiment 2, NT embryos were treated with 0.5 nM PCI-24781 or 10 nM quisinostat for different durations. The rate of blastocyst formation was significantly higher in the 0.5 nM PCI-24781 group than in the control (25.3 v. 10.5%; P < 0.05; Table 1). Moreover, treatment with 10 nM quisinostat dramatically increased the proportion of embryos that reached the blastocyst stage, in comparison with the control group (18.4 v. 10.7%; P < 0.05). Table 1 shows that SCNT embryos treated with 0.5 nM PCI-24781 for 6 h had higher rates of blastocyst formation than control group (25.2 v. 10.2%; P < 0.05). However, the rate of blastocyst formation was significantly higher in the 10 nM quisinostat for 24 h group than in the control (19.8 v. 10.1%; P < 0.05). We determined the treatment conditions of PCI-24781 (0.5 nM for 6 h) and quisinostat (10 nM for 24 h) that significantly improve the in vitro development of pig SCNT embryos.
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