Lee Seok Hee, Oh Hyun Ju, Kim Min Jung, Setyawan Erif M N, Choi Yoo Bin, Lee Byeong Chun
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
Mol Reprod Dev. 2018 Apr;85(4):336-347. doi: 10.1002/mrd.22969. Epub 2018 Mar 23.
Human endothelial progenitor cells (EPCs) have been applied to regenerative medicine for their roles in angiogenesis as well as neovascularization, and these angiogenetic functions have beneficial effects on maturation of ovarian follicles. However, little information is available on whether EPCs on culture systems affect oocyte maturation and subsequent embryo development. Therefore, the objective of this study was to investigate the effect of EPC co-culture on porcine oocytes during in vitro maturation (IVM) and subsequent embryo development, and to examine gene expression in cumulus cells, oocytes and blastocysts. The effect of co-culture using EPC on porcine oocyte IVM was investigated. Oocytes were activated using electrical stimulation and embryo developmental competence was estimated. The expression of the genes related to cumulus expansion, oocyte maturation, embryo development, and apoptosis were analyzed. In result, there was a significantly increased maturation rate in EPC group compared with control (p < 0.05). Also, oocytes co-cultured with EPCs exhibited significantly improved blastocyst formation rates (p < 0.05). The expression of mRNAs associated with cumulus expansion and apoptosis in cumulus cells was significantly up-regulated in EPC group. Also, markedly increased levels of GDF9, BMP15, and BCL2 were observed in oocytes from the EPC group. Blastocysts in the co-culture group showed significantly higher SOX2, OCT4, and NANOG levels. In conclusion, co-culturing porcine oocytes with EPCs improves their maturation by regulating genes involved in cumulus cell expansion, oocyte maturation, and apoptosis. Moreover, EPC co-culture during IVM enhanced embryo development as shown by increased blastocyst formation rate and pluripotency-related gene expression.
人类内皮祖细胞（EPCs）因其在血管生成和新血管形成中的作用而被应用于再生医学，这些血管生成功能对卵巢卵泡的成熟具有有益影响。然而，关于培养系统中的EPCs是否影响卵母细胞成熟和随后的胚胎发育，目前所知甚少。因此，本研究的目的是探讨EPC共培养对猪卵母细胞体外成熟（IVM）及随后胚胎发育的影响，并检测卵丘细胞、卵母细胞和囊胚中的基因表达。研究了使用EPC进行共培养对猪卵母细胞IVM的影响。使用电刺激激活卵母细胞并评估胚胎发育能力。分析了与卵丘扩展、卵母细胞成熟、胚胎发育和细胞凋亡相关的基因表达。结果显示，与对照组相比，EPC组的成熟率显著提高（p < 0.05）。此外，与EPCs共培养的卵母细胞的囊胚形成率显著提高（p < 0.05）。EPC组卵丘细胞中与卵丘扩展和细胞凋亡相关的mRNA表达显著上调。此外，在EPC组的卵母细胞中观察到GDF9、BMP15和BCL2水平明显升高。共培养组的囊胚显示SOX2、OCT4和NANOG水平显著更高。总之，将猪卵母细胞与EPCs共培养可通过调节参与卵丘细胞扩展、卵母细胞成熟和细胞凋亡的基因来提高其成熟度。此外，IVM期间的EPC共培养增强了胚胎发育，表现为囊胚形成率增加和多能性相关基因表达增加。
