Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Infertility and Reproductive Health Research Center (IRHRC), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Cell Tissue Bank. 2021 Mar;22(1):103-114. doi: 10.1007/s10561-020-09870-z. Epub 2020 Oct 9.
Vitrification of embryos has been known as the most efficient cryopreservation method in assisted reproductive technology clinics. Vitrification of preimplantation embryo might be associated with altered gene expression profile and biochemical changes of vitrified embryos. Stringent regulation of gene expression in early embryonic stages is very critical for normal development. In the present study, we investigated the effect of vitrification on the canonical miRNA biogenesis pathway, and also the expression of developmental related miRNAs, in 8-cell and blastocyst mouse embryos. Although the expression pattern of the miRNA biogenesis pathway genes differed between 8-cell and blastocyst mouse embryos, vitrification did not affect the expression level of these genes in preimplantation embryos. The expression levels of miR-21 and let-7a were significantly decreased in vitrified 8-cell embryos and fresh blastocysts when compared with fresh 8-cell embryos. The expression of Stat3 was significantly reduced in blastocysts after vitrification. The alteration in the expression pattern of miRNAs, due to their mode of action, can affect broad downstream key developmental signaling pathways. Therefore, the blastocyst stage is the preferred point for embryo vitrification as they are less susceptible to cryo-damages regarding the stability of miRNAs related to the developmental and implantation competence of embryo.
胚胎玻璃化已被认为是辅助生殖技术临床中最有效的冷冻保存方法。胚胎玻璃化可能与冷冻胚胎的基因表达谱改变和生化变化有关。早期胚胎严格的基因表达调控对正常发育至关重要。在本研究中,我们研究了玻璃化对 8 细胞期和囊胚期小鼠胚胎中经典 miRNA 生物发生途径以及发育相关 miRNA 表达的影响。尽管 miRNA 生物发生途径基因的表达模式在 8 细胞期和囊胚期小鼠胚胎之间存在差异,但玻璃化并未影响植入前胚胎中这些基因的表达水平。与新鲜的 8 细胞胚胎相比,玻璃化的 8 细胞胚胎和新鲜囊胚中 miR-21 和 let-7a 的表达水平显著降低。玻璃化后囊胚中 Stat3 的表达显著降低。由于 miRNA 的作用模式,其表达模式的改变可能会影响广泛的下游关键发育信号通路。因此,囊胚阶段是胚胎玻璃化的首选阶段,因为它们在与胚胎发育和植入能力相关的 miRNA 稳定性方面对冷冻损伤的敏感性较低。