Studies On Long-Tailed Macaque (Macaca Fascicularis)-Pig Interspecies Somatic Cell Nuclear Transfer

The primary objective of long-tailed macaque-pig interspecies somatic cell nuclear transfer (iSCNT) was to yield embryo stem cells (ESCs) from iSCNT embryos which had similar genetic background as human. Then it would become the material for medical and science researches. iSCNT research wouldn't only support basic researches, such as human therapeutic cloning, developmemt of early embryo in primate, and nuclear-cytoplasm interactions, nuclear reprogramming and epigenetic during nuclear transfer, but would also provide reference for iSCNT in restoring endangered, or even extinct species and populations. The main contents and results were as following:I:To investigate the effects of different fusion/activation parameters on iSCNT embryos development for selecting appropriate fusion/activation parameter in iSCNT, and the development of iSCNT embryos in different embryo mediums for selecting appropriate embryo medium for iSCNT embryos. The results suggested that when long tailed macaque ear fibroblasts (MEF) with diameter 20μm, as donor cells, were introduced into pig nucleated oocytes, electrical pulses (60 V/mm or 80 V/mm,30μs,2 direct current pluses, with 1.0 s interval) were applied to fuse/activate the iSCNT embryos to obtain the highest fusion/activation rate. The more important, the resulting iSCNT embryos could develop into blastocyst stage. Then, one set of electrical pulses (80 V/mm,30μs,2 direct current, with 1.0 s interval) was applied to accomplish the fusion/activation of iSCNT embryos. Both PZM-3 and HECM-10 supported the development of iSCNT embryos.II:To investigate the effects of two histone deacetylase inhibitors (HADCi), Trichostatin A (TSA) and Valproic acid (VPA), on the development of iSCNT embryos. The results suggested that when PZM-3 as embryo medium, 10 nM TSA treatment for 48 h incresed blastocyst rate significantly (P<0.05)...