Wang Xiaohan, Lu Fangting, Bai Shun, Wu Limin, Huang Lingli, Zhou Naru, Xu Bo, Wan Yangyang, Jin Rentao, Jiang Xiaohua, Tong Xianhong
Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Division of Life Sciences and Medicine, Reproductive and Genetic Hospital, The First Affiliated Hospital of USTC, University of Science and Technology of China, Hefei, China.
Front Genet. 2022 Jan 28;12:815270. doi: 10.3389/fgene.2021.815270. eCollection 2021.
Human autologous sperm freezing involves ejaculated sperm, and testicular or epididymal puncture sperm freezing, and autologous sperm freezing is widely used in assisted reproductive technology. In previous studies, researchers have tried to cryopreserve sperm from mammals (rats, dogs, etc.) using a -80°C freezer and have achieved success. It is common to use liquid nitrogen vapor rapid freezing to cryopreserve human autologous sperm. However, the operation of this cooling method is complicated, and the temperature drop is unstable. In this study, we compared the quality of human ejaculation and testicular sperm after liquid nitrogen vapor rapid freezing and -80°C freezing for the first time. By analyzing sperm quality parameters of 93 ejaculated sperm and 10 testicular sperm after liquid nitrogen vapor rapid freezing and -80°C freezing, we found reactive oxygen species (ROS) of sperm of the -80°C freezer was significantly lower than liquid nitrogen vapor rapid freezing. Regression analysis showed that progressive motility, ROS, and DNA fragmentation index (DFI) in post-thaw spermatozoa were correlated with sperm progressive motility, ROS, and DFI before freezing. For the freezing method, the -80°C freezer was positively correlated with the sperm progressive motility. Among the factors of freezing time, long-term freezing was negatively correlated with sperm progressive motility and ROS. Although freezing directly at -80°C freezer had a slower temperature drop than liquid nitrogen vapor rapid freezing over the same period, the curves of the temperature drop were similar, and slight differences in the freezing point were observed. Furthermore, there were no statistically significant differences between the two methods for freezing testicular sperm. The method of direct -80°C freezing could be considered a simplified alternative to vapor freezing for short-term human sperm storage. It could be used for cryopreservation of autologous sperm (especially testicular sperm) by fertilization centers. Clinical Trial Registration: (website), identifier (ChiCTR2100050190).
人类自体精子冷冻包括射出精子、睾丸或附睾穿刺精子冷冻，自体精子冷冻在辅助生殖技术中被广泛应用。在以往研究中，研究人员尝试使用-80℃冰箱对哺乳动物（大鼠、狗等）的精子进行冷冻保存并取得了成功。使用液氮蒸汽快速冷冻来保存人类自体精子很常见。然而，这种冷却方法操作复杂，温度下降不稳定。在本研究中，我们首次比较了液氮蒸汽快速冷冻和-80℃冷冻后人类射出精子和睾丸精子的质量。通过分析93例射出精子和10例睾丸精子在液氮蒸汽快速冷冻和-80℃冷冻后的精子质量参数，我们发现-80℃冰箱保存的精子活性氧（ROS）显著低于液氮蒸汽快速冷冻保存的精子。回归分析表明，解冻后精子的前向运动能力、ROS和DNA碎片指数（DFI）与冷冻前精子的前向运动能力、ROS和DFI相关。对于冷冻方法，-80℃冰箱与精子前向运动能力呈正相关。在冷冻时间因素中，长期冷冻与精子前向运动能力和ROS呈负相关。虽然在同一时期直接在-80℃冰箱冷冻的降温速度比液氮蒸汽快速冷冻慢，但降温曲线相似，且在冰点观察到细微差异。此外，两种冷冻睾丸精子的方法之间没有统计学上的显著差异。直接-80℃冷冻方法可被视为一种简化的替代方法，用于短期人类精子储存的蒸汽冷冻。它可被受精中心用于自体精子（尤其是睾丸精子）的冷冻保存。临床试验注册：（网站），标识符（ChiCTR2100050190）。
