Department of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
Department of Animal Health and Anatomy, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
Int J Mol Sci. 2020 Sep 25;21(19):7067. doi: 10.3390/ijms21197067.
Oocyte cryopreservation has a significant impact on subsequent embryonic development. Herein, we investigated whether supplementing in vitro maturation medium with Leukemia Inhibitory Factor (LIF) prior to vitrification affects embryo development and gene expression at different embryo developmental stages. A panel of genes including maternal effect, epigenetics, apoptosis and heat stress was relatively quantified. The results show reduced cleavage rates after vitrification, regardless of the LIF treatment. Although not statistically different from control-vitrified oocytes, oocyte apoptosis and the blastocyst yield of LIF-vitrified oocytes were similar to their non-vitrified counterparts. Vitrification increased oocyte , and gene expression while its expression decreased in LIF-vitrified oocytes to similar or close levels to those of non-vitrified oocytes. With a few gene-specific exceptions, vitrification significantly increased the expression of , , , and in oocytes and most stages of embryo development, while comparable expression patterns for these genes were observed between LIF-vitrified and non-vitrified groups. Vitrification increased expression in oocytes and in 2-cell embryos. Our data suggest that vitrification triggers stage-specific changes in gene expression throughout embryonic development. However, the inclusion of LIF in the IVM medium prior to vitrification stimulates blastocyst development and several other developmental parameters and induces oocytes and embryos to demonstrate gene expression patterns similar to those derived from non-vitrified oocytes.
卵母细胞冷冻保存对后续胚胎发育有重大影响。在此,我们研究了在玻璃化之前向体外成熟培养基中添加白血病抑制因子 (LIF) 是否会影响不同胚胎发育阶段的胚胎发育和基因表达。相对定量了包括母体效应、表观遗传学、细胞凋亡和热应激在内的一组基因。结果显示,无论是否进行 LIF 处理,玻璃化后卵裂率均降低。尽管与对照玻璃化卵母细胞相比没有统计学差异,但 LIF 玻璃化卵母细胞的卵母细胞凋亡和囊胚产量与未玻璃化卵母细胞相似。玻璃化增加了卵母细胞和胚胎发育各个阶段的 、 和 基因表达,而 LIF 玻璃化卵母细胞中的表达则降低到与未玻璃化卵母细胞相似或接近的水平。除了少数基因特异性例外,玻璃化显著增加了卵母细胞和胚胎发育各个阶段的 、 、 、 和 基因的表达,而 LIF 玻璃化和未玻璃化组之间观察到这些基因的表达模式相似。玻璃化增加了卵母细胞中的 表达和 2 细胞胚胎中的 表达。我们的数据表明,玻璃化在整个胚胎发育过程中引发了特定阶段的基因表达变化。然而,在玻璃化之前将 LIF 纳入 IVM 培养基中会刺激囊胚发育和其他几个发育参数,并诱导卵母细胞和胚胎表现出与未玻璃化卵母细胞相似的基因表达模式。