| 其他摘要 | While the vast majority of animal research is conducted in rodents, we believe that the unanswered questions regarding ESCs in vivo function, apoptosis, and tumor formation would be best addressed using a non-human primate (NHP). Rhesus macaques (the standard nonhuman primate model organism) possess remarkable anatomical, physiological, and metabolic similarities to humans, and many human neurological diseases, such as Alzheimer and Parkinson diseases, can only be accurately modeled in the nonhuman primate. In this thesis, the mechanism of maintaining self-renewal and pluripotency of embryonic stem cells has been reviewed, and some research findings have been made in the following: 1) We applied the oligonucleotide array and quantitative real-time PCR (qPCR) to analyze the expression profiles of the five rhesus fibroblast cell lines, in the hope of finding out the differentially expressed genes between the supportive and non-supportive feeder cell lines. We investigated the cytokines located in the extracellular region or on the membrane of fibroblast cells, which may interact with rESCs through membrane-binding receptors and trigger the downstream signaling events that finally prolonged the undifferentiated growth of rESCs. Eight genes, GREM2, bFGF, KITLG, DKK3, GREM1, AREG, SERPINF1 and LTBP1, were found to be overexpressed in supportive feeder cells, and three of them, SCF, bFGF and GREM2, were validated by qPCR. 2) In order to depict the WNT expression patterns in IVF, ICSI (intracytoplasmic sperm injection), SCNT and parthenogenetic blastocysts and to figure out whether there are differences between IVF and SCNT blastocysts and between IVF and parthenogenetic blastocysts, we combined the pathway-specific PCR Array System and immunocytochemistry to monitor the WNT expression in both mRNA and protein level. We also tested the WNT signaling pathway components in ICSI bastocysts with the aim of eliminating the effect of micromanipulation. Noncanonical WNT/JNK pathway, instead of canonical WNT pathway, may be active in rhesus IVF blastocysts. Moreover, expression of WNT signaling pathway genes in SCNT and parthenogenetic embryos are significantly higher than those in IVF embryos. |
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