The rhesus monkey has been one of the most widely used laboratory animal in medical and biological research due to the close relationship to human being in anatomy, genetics and physiology. Rhesus monkey embryonic stem cell (rESC) is the ideal model for study on embryologic developmental biology and cell replacement therapy for human cellular degenerative diseases. Meanwhile, to obtain sufficient oocytes by superovulation (SO) is essential process for embryonic biotechnology, and embryo transfer (ET) is an important way to produce offspring from in vitro produced embryos. In this thesis, the progress in neural differenatioation from primate ESCs and assisted reproductive technology (ART) in primate were reviewed. Furthermore, experiments focused on neural differentiation from rESCs and the time windows during SO-ET in rhesus monkeys were designed and ferformed, which including: 1) The p53 and its regulator ASPP family, p53 targets Bax, PIG-3 and p21waf1 were detected during the process of neural differentiation from rESCs. The results showed that FGF-2 and other neural induction factors promote cell proliferation, neural conversion, and apoptosis during the differentiation process.The ASPP family members changed their expression either in quantity or pattern to regulate p53 in this process. The p53 actived its proapoptotic targets Bax and PIG-3 but not cell cycle arrest target p21waf1. The results implied that neural differentiation is concomitant with apoptosis in rESCs. 2) The roles of folic acid (FA) during neural differentiation from rESCs were assayed by regulating FA concentration or introducing the antagonist of FA: methotrexate (MTX). Our data showed that FA deficiency decreased cell proliferation and reduced the formation of neural rosettes. No obvious difference in expression of nestin and Pax6 (neural progenitor markers) among neural progenitors obtained from all FA concentration groups was observed. However, FA deficiency affected neuronal differentistion, but not neuroglia. Furtherly, MTX reduced cell expansion, neural induction and neuronal differnatiation, and excessive FA supply was able to partly rescue MTX’s effects. 3）The results of SO and ET in rhesus monkeys were summarized and the reasons that could result in SO-ET failure were analyzed. We found that the time windows such as age stages, repeated stimulating with recombinant human gonadotropins and restimulation interval, and breeding season were the key factors for SO-ET. Importantly, 2-8-cell embryos to blastocysts were transferred through oviduct embryo transfer. We found that the optimal time window for embryo transfer in rhesus monkeys might be the stage from late follicular phase to early luteal phase. For the first time, we found some novel roles of p53 and folic acid during the rESCs neural differentiation. These preliminary data were expected to be helpful for learning correlative mechanism of normal or abnormal development in nervous system. And the finding of time window for embryo transfer might be an important reference to further work.
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