Grupo de Investigación en Biodiversidad y Genética Molecular (BIOGEM), Departamento de Producción Animal, Universidad Nacional de Colombia, Sede Medellín, Carrera 65 No. 59 A-110, Código, 050034, Colombia.
Grupo de Investigación en Biodiversidad y Genética Molecular (BIOGEM), Departamento de Producción Animal, Universidad Nacional de Colombia, Sede Medellín, Carrera 65 No. 59 A-110, Código, 050034, Colombia.
Cryobiology. 2019 Aug;89:42-50. doi: 10.1016/j.cryobiol.2019.05.008. Epub 2019 May 16.
Although vitrification is the current routine method for human embryo cryopreservation, it may cause detrimental effects. The aim of this study was to evaluate the effect of supplementing in vitro culture (IVC) media and/or vitrification solutions (VS) with Resveratrol on the presence of apoptotic markers, reactive oxygen species (ROS) level, glutathione (GSH) content and relative gene abundance. Abattoir-derived oocytes were matured and fertilized in vitro according to a standard procedure. Zygotes were cultured in IVC medium supplemented with or without 0.5 μM Resveratrol (C, C respectively). On day 7, blastocysts were vitrified using the minimum volume vitrification method supplementing VS with (CV, CV) or without (CV, CV) 0.5 μM Resveratrol. After warming, embryonic quality parameters were evaluated. Survival rates were significantly lower in CV group compared with CV group, but no differences in hatching rate were observed between groups. Vitrification/warming process did not alter total cell number or the presence of apoptotic or dead cells, but CV and CV groups presented a significant increase in dead cells (P < 0.05 by ANOVA). Resveratrol supplementation in VS (CV) restored GSH content (P < 0.05) to the level found in the C group. Vitrification/warming process significantly increased the expression of FOXO3A, PNPLA2, BCL2L1 and BAX genes (P < 0.05). Resveratrol addition to IVC medium or VS partially compensated this increase for FOXO3A and PNPLA2 (P < 0.05) but not for BCL2L1 and BAX. In conclusion, supplementation of IVC media or VS with 0.5 μM resveratrol may help embryos to partially restore the initial quality they had before the cryopreservation process.
尽管玻璃化法是目前人类胚胎冷冻保存的常规方法,但它可能会造成有害影响。本研究旨在评估在体外培养(IVC)培养基和/或玻璃化溶液(VS)中添加白藜芦醇对凋亡标志物、活性氧(ROS)水平、谷胱甘肽(GSH)含量和相对基因丰度的影响。屠宰场来源的卵母细胞按照标准程序体外成熟和受精。将胚胎在添加或不添加 0.5μM 白藜芦醇(C、C 组)的 IVC 培养基中培养。第 7 天,使用最小体积玻璃化法将胚胎玻璃化,VS 中添加(CV、CV)或不添加(CV、CV)0.5μM 白藜芦醇。解冻后,评估胚胎质量参数。与 CV 组相比,CV 组的存活率显著降低,但两组的孵化率无差异。玻璃化/解冻过程并未改变总细胞数或凋亡或死亡细胞的存在,但 CV 和 CV 组的死亡细胞数量显著增加(ANOVA 分析,P<0.05)。VS 中添加白藜芦醇(CV)使 GSH 含量恢复到 C 组的水平(P<0.05)。玻璃化/解冻过程显著增加了 FOXO3A、PNPLA2、BCL2L1 和 BAX 基因的表达(P<0.05)。在 IVC 培养基或 VS 中添加 0.5μM 白藜芦醇部分补偿了 FOXO3A 和 PNPLA2 的这种增加(P<0.05),但对 BCL2L1 和 BAX 没有影响。总之,在 IVC 培养基或 VS 中添加 0.5μM 白藜芦醇可能有助于胚胎部分恢复冷冻保存过程前的初始质量。