人体外成熟卵母细胞的玻璃化:对线粒体超微结构和耗氧量的影响

Shubiao Han, W. Han, Xiaodong Zhang, Junxia Liu, G. Huang
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摘要

背景:本研究旨在评估玻璃化对人IVM卵母细胞线粒体的影响。方法:从卵巢刺激周期获得401个未成熟卵母细胞,IVM后随机分为新鲜组和玻璃化组。根据解冻后的培养时间,将玻璃化组分为0小时(0 h)、2小时(2 h)和4小时(4 h)亚组。比较四组小鼠线粒体形态和耗氧量。计算ICSI受精后的正常受精率、卵裂率和囊胚形成率。结果:与对照组相比,0 h和2 h组线粒体结构平均灰度值显著降低(分别为0.48±0.09、0.50±0.36、0.61±0.12);P[公式:见文]0.05),4 h组恢复率为(0.61±0.24∶0.61±0.12,P[公式:见文]0.05)。此外,与新鲜组相比,0 h和2 h组的耗氧量也显著降低(分别为2.91±0.77 fmol/s、3.26±1.34 fmol/s和3.96±1.44 fmol/s);P[公式:见文]0.05),培养4 h后恢复(分别为3.96±1.44 fmol/s和4.41±1.38 fmol/s);P[公式:见正文]0.05)。正常受精率和卵裂胚率在4组间无显著差异,但囊胚发育率在0 h组显著降低。结论:这些结果表明,在玻璃化过程中,卵母细胞的耗氧量和线粒体结构可能会发生暂时的动态变化,但在4小时后出现恢复。
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Vitrification of Human In-Vitro Matured Oocytes: Effects on Mitochondrial Ultrastructure and Oxygen Consumption
Background: This study was conducted to evaluate the impact of vitrification on mitochondrial of human IVM oocytes. Methods: A total of 401 immature oocytes were obtained from ovarian stimulated cycles, which were randomly divided into fresh and vitrification groups after IVM. According to the cultured time after thawing, the vitrification groups were divided into 0 hours (0 h), 2 hours (2 h), or 4 hours (4 h) subgroups. Mitochondrial morphology and oxygen consumption were compared among the four groups. After fertilization by ICSI, normal fertilization, cleaved embryos, and blastocyst formation rate were also calculated. Results: The mean gray value of mitochondria structure was significantly decreased in 0 h and 2 h groups when compared to control group (0.48 ± 0.09, 0.50 ± 0.36 vs. 0.61 ± 0.12, respectively; P [Formula: see text] 0.05), and recovered (0.61 ± 0.24 vs. 0.61 ± 0.12, P [Formula: see text] 0.05) in 4 h group. In addition, oxygen consumption was also significantly decreased in 0 h and 2 h groups compared to fresh (2.91 ± 0.77 fmol/s, 3.26 ± 1.34 fmol/s vs. 3.96 ± 1.44 fmol/s, respectively; P [Formula: see text] 0.05), and recovered after 4 h culture (3.96 ± 1.44 fmol/s vs. 4.41 ± 1.38 fmol/s, respectively; P [Formula: see text] 0.05). The percentage of normal fertilization and cleaved embryos were no differences among the four groups, however, blastocyst development rate was significantly lower in 0 h group. Conclusion: These results indicate that during the vitrification process, the oxygen consumption and mitochondrial structure of oocytes may undergo temporary dynamic changes, but appear to recover by 4 hours.
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审稿时长
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