胎牛血清在猪孤雌囊胚玻璃化冷冻后恢复培养中的作用研究

【目的】研究胎牛血清(fetal bovine serum, FBS)在猪孤雌囊胚玻璃化冷冻后恢复培养中的作用。【方法】本试验以体外培养第5天的猪孤雌激活囊胚为材料，将新鲜和冷冻囊胚分别在含10%FBS(V/V)的胚胎培养液中继续培养48 h,即分为新鲜组(Fresh)、新鲜+FBS组(Fresh+FBS)、冷冻组(Vitrified)、冷冻+FBS组(Vitrified+FBS)。观察各组囊胚的扩张和孵化能力，检测胚胎的细胞膜损伤、凋亡细胞数目、总细胞数目、胞内活性氧(ROS)水平、线粒体活性以及发育相关基因的表达水平。【结果】与Fresh和Vitrified组相比，Fresh+FBS和Vitrified+FBS组的完全扩张率、孵化率和囊胚细胞总数均显著提高(P<0.05),细胞膜损伤率和细胞凋亡率均显著降低(P<0.05)。与Fresh组相比，Vitrified组ROS水平显著升高(P<0.05),Fresh+FBS和Vitrified+FBS组ROS水平均显著降低(P<0.05)。Vitrified+FBS组的线粒体活性显著高于Vitrified组(P&l...

The Study on the Role of Fetal Bovine Serum Supplementation During Postwarming Culture on Vitrified Porcine Parthenogenetic Blastocysts

【Objective】 The aim of this study was to evaluate the effects of fetal bovine serum (FBS) supplementation during postwarming culture on vitrified porcine parthenogenetic blastocysts.【Method】 The porcine parthenogenetic blastocysts on the 5th day of in vitro culture were used in this experiment,and the fresh and vitrified blastocysts were cultured in the embryo culture medium containing 10% FBS (V/V) for 48 h.This experiment was divided into four groups:Fresh,Fresh+FBS,Vitrified and Vitrified+FBS groups.The blastocysts expansion and hatching rates,the percentages of membrane damage and apoptosis and total cell number were determined.Moreover,the intracellular reactive oxygen species (ROS) levels,mitochondrial activity and expression levels of genes related embryo development in the blastocysts were detected.【Result】 Compared with the Fresh and Vitrified groups, the complete expansion rate, hatching rate and total number of blastocyst cells in the Fresh+FBS and Vitrified+FBS groups were significantly increased (P<0.05), while the cell membrane damage rate and apoptotic cell rate were significantly decreased (P<0.05). Compared with the Fresh group, ROS levels were significantly increased in the Vitrified group, while ROS levels were significantly decreased in both the Fresh+FBS and Vitrified+FBS groups (P<0.05). The mitochondrial activity of Vitrified+FBS group was significantly higher than that of Vitrified+FBS group and significantly lower than that of Fresh+FBS group (P<0.05), but there was no significant difference between Vitrified+FBS group and Fresh group (P>0.05). Compared with the Fresh group, POU class 5 homeobox 1(POU5F1) gene expression level in Vitrified group was significantly increased (P<0.05) and catalase (CAT)gene expression level was significantly decreased (P<0.05). There were no significant differences in the expression levels of proliferating cell nuclear antigen (PCNA), DNA methyltransferase 3A (DNMT3A), superoxide dismutase 1 (SOD1) and BCL2-associated X protein (BAX):BCL2L1 between the Fresh and Vitrified groups (P>0.05). The expression levels of PCNA, SOD1 and CAT genes in Fresh+FBS and Vitrified+FBS groups were significantly higher than those in Fresh and Vitrified+FBS groups (P<0.05).【Conclusion】 This study found that fresh and vitrified parthenogenes blastocysts were cultured in 10% FBS on the 5th day of in vitro culture, and their development ability and quality of embryo were significant improved.