Post Graduate School of Universitas Airlangga Surabaya, Indonesia.
Department of Biomedical Science, Faculty of Medicine Universitas Airlangga Surabaya, Indonesia.
Pol J Vet Sci. 2022 Mar;25(1):19-26. doi: 10.24425/pjvs.2022.140836.
The process of vitrification of the cumulus-oocyte complex (COCs) often results in cold shock. When warming, heat shock occurs which can disrupt the balance of intracellular calcium (Ca2+) intensity. Drastic changes in temperature cause Reactive Oxygen Species (ROS), affecting changes on Ca2+ in COCs. The role of calcium is needed for oocyte activation in the fertilization process. The purpose of this study was to measure the expression of Ca2+ and the intensity of Ca2+ in COCs after vitrification. The study was divided into 2 groups, the control group (C) of fresh COCs, and the treatment group (T) of COCs after vitrification. After vitrification for 24 hours, then thawing, the expression of Ca2+ was examined using the Immunocytochemistry (ICC) method and the intensity of calcium (Ca2+) with a Confocal Laser Scanning Microscope (CLSM). The research data obtained were analyzed statistically by T-Test. The results showed that the expression of Ca2+ in the control group (12.00±0.00) was different from the treatment group (0.35±0.79). The intensity of Ca2+ in the control group (1059.43±489.59) was different from the treatment group (568.21±84.31). The conclusion of this study is that cryopreservation affects calcium in COCs; there were differences in the expression and the intensity of Ca2+ between fresh COCs and COCs after vitrification. Ca2+ intensity of COCs after vitrification was concentrated in the nucleus, while in fresh COCs it was concentrated in the cytoplasm.
卵丘-卵母细胞复合物（COCs）的玻璃化过程通常会导致冷休克。当解冻时，会发生热休克，从而破坏细胞内钙（Ca2+）强度的平衡。温度的剧烈变化会导致活性氧（ROS），从而影响 COCs 中的 Ca2+变化。钙在卵母细胞受精过程中的激活中起着重要作用。本研究的目的是测量玻璃化后 COCs 中 Ca2+的表达和 Ca2+强度。研究分为新鲜 COCs 的对照组（C）和玻璃化 COCs 的处理组（T）两组。玻璃化 24 小时后，然后进行解冻，使用免疫细胞化学（ICC）方法检查 Ca2+的表达，并使用共聚焦激光扫描显微镜（CLSM）检查 Ca2+强度。通过 T 检验对研究数据进行统计分析。结果表明，对照组（12.00±0.00）的 Ca2+表达与处理组（0.35±0.79）不同。对照组（1059.43±489.59）的 Ca2+强度与处理组（568.21±84.31）不同。本研究的结论是，冷冻保存会影响 COCs 中的钙；新鲜 COCs 和玻璃化 COCs 之间在 Ca2+的表达和强度方面存在差异。玻璃化 COCs 的 Ca2+强度集中在核内，而新鲜 COCs 的 Ca2+强度集中在细胞质中。
