Buffalo-pig Inter-species Somatic Cell Nuclear Transfer

Inter-species somatic cell nuclear transfer (iSCNT), namely transplant a somatic cell into a distant species oocyte, has been used in many fields including medicine science, saving endangered animals and as a tool to study the nuclear reprogramming. There’re numerous factors influence the development of buffalo-pig iSCNT. The aim of this present study is to optimize the electronic fusion/activation voltage, the culture conditions of the ratio of embryo number/medium volume, as well as detect the efficiency of chemical activation using ionomycin plus6-DMAP. In addition, buffalo fibroblasts labeled by GFP reporter gene as nuclear transfer donors used to examine the reprogramming efficiency of porcine oocytes. The results are listed as follows:1. Different electronic fusion/activation voltage (120,150and180v/mm) does not significantly affect the in vitro development of buffalo-pig iSCNT embryos, whereas the150v/mm generated a better result.2. Electrical fusion/activation, electrical fusion/activation followed by chemical activation using ionomycin plus6-DMAP have been performed to compare their efficiency. Data indicated no difference among two methods when the cleavage and blastocyst rates had been calculated.3. We detected the optimal ratio of embryo number/medium volume for cultivating of iSCNT embryos, and results indicates that this has no effect on the buffalo-pig development.4. Pig oocytes are capable of reprogramming the buffalo somatic nuclei. The iSCNT embryos can develop to blastcoyst stage in vitro and express exogenous GFP during the whole developmental stage.In conclusion, we optimized the electronic fusion/activation voltage, the culture conditions of the ratio of embryo number/medium volume, as well as detect the efficiency of chemical activation using ionomycin plus6-DMAP. The expression of exogenous GFP in iSCNT embryos suggesting the pig oocytes are capable of reprogramming the buffalo somatic nuclei.