In vitro production of cloned and transgenically cloned embryos from Guangxi Huanjiang Xiang pig.

Guangxi Huanjiang Xiang pig is a unique miniature pig strain that is originally from Huanjiang Maonan Autonomous County of Guangxi province, China, and shows great potential in agricultural and biomedical research. Although cloning and genetic modification of this pig would enhance its application value, cloning of this strain has not yet been reported. We sought to establish appropriate cloning procedures and produce transgenic embryos in Huanjiang Xiang pigs through the following methods. We isolated fibroblasts from tails of Huanjiang Xiang pig and genetically modified them using Xfect transfection. Fibroblasts, either in non-transgenic or transgenic forms, were used as donor cells for reconstructed embryos by somatic cell nuclear transfer (SCNT), and in vitro development was monitored after the reconstruction. We found no difference in blastocyst formation rate between non-transgenic and transgenic embryos (10.8% vs. 10.3%; P ≥ 0.05). In addition, we tested whether Scriptaid, a widely used histone deacetylase inhibitor, could enhance the in vitro development of Huanjiang Xiang pig cloned embryos. Treatment with 500 nM Scriptaid for 16 h post-activation significantly increased the blastocyst formation rate (26.1% vs. 10.8% for non-transgenic nuclear transfer groups with vs. without the Scriptaid treatment and 28.5% vs. 10.3% for transgenic nuclear transfer groups with vs. without the Scriptaid treatment; P < 0.05). This study provided a basis for further generation of cloned and transgenically cloned Huanjiang Xiang pigs used in agricultural and biomedical research.