| There are lots of advantages for pigs in the research on human medicine and basicresearch, therefore, much endeavors were placed on the porcine somatic cell nucleartransfer (SCNT). However, the efficiency of porcine SCNT was still very low. Themajor reason maybe lies on the incomplete or faulty epigenetic reprogramming indifferentiated donor cells. Recently, several deacetylase inhibitors were applied on theporcine SCNT embryos culturing, and most of them showed a positive effect on theimproving of in vivo developmental competency. In addition, recloning waspreviously taken as another effective mean in the promoting the efficiency ofreprogramming. However, the recloning efficiency was varied in different species andit was still low in pig. In the present study, vaproic acid (VPA) was used to the studyon the improvement of porcine recloned embryos. The HDACi valproic acid is acell-permeable, short-chain fatty acid, which has been shown to play a significant rolein the reprogramming of mouse fibroblasts.In the present study, the cloned fetal fibroblast cells were successfully isolated.Following, the growth characteristics on proliferation rate, cell cycle and apoptosisbetween cloned fetal fibroblast cells and normal fetal fibroblast cells were analyzed.However, there were no significant differences on the growth characteristics betweenthese two kinds of cells. The best treated time and concentration were then analyzedon the in vitro development of recloned embryos. The results showed that a highblastocyst-formation rate was reached when1mmol/L VPA was used to reclonedembryos treatment for16h following activation. The level of acH3K9was alsoexamined, and it showed that acH3K9was high in VPA treated recloned embryos.Through quantitative RT-PCR, the results revealed that there were no significantdifferences on the expression level of Oct4、Nanog、Sox2and Klf4.In conclusion, the in vitro developmental capability of recloned embryos could beimproved following VPA treatment. |
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