作者：彭飞宇¹、刘理想¹、杨镒峰、张旭林、张燠诗、范冰峰、褚萧宇、曹杰、张颖、赵伟刚、唐毓^*、许保增^*

刊物名称：Cryobiology

发表年月：2026.02.18

摘要内容：Sperm cryopreservation imposes osmotic stress due to the addition and removal of high-concentration cryoprotectants. Whether organic osmolytes can alleviate this stress to improve cryopreservation efficiency remains unclear. Here, we evaluated the effects of glycine and betaine, supplemented as organic osmolytes throughout the cryopreservation process, on the quality and developmental potential of cryopreserved sika deer sperm. Pooled semen samples from five healthy adult stags were randomly allocated to four groups: a control (base extender) and three treatment groups supplemented with glycine and betaine at 6 mM + 200 μM, 10 mM + 500 μM, and 14 mM + 800 μM, respectively. The addition of 10 mM glycine and 500 μM betaine significantly improved post-thaw sperm quality, including enhanced total motility (TM), progressive motility (PM), average path velocity (VAP), curvilinear velocity (VCL), plasma membrane and acrosome integrity, and fertilization capacity, while reducing reactive oxygen species (ROS) levels compared with the control and other treatments. These findings suggest that glycine and betaine at 10 mM and 500 μM, respectively, may mitigate cryo-injury and enhance cryosurvival in sika deer sperm, offering a promising strategy for improving semen cryopreservation and artificial insemination protocols

Figure 2. Glycine and betaine protect sperm membrane and acrosome integrity after thawing. (A) Detection of sperm plasma membrane integrity by HOST (scale, 50 μm). a: sperm with intact plasma membrane, b: sperm with damaged plasma membrane; (B) Sperm plasma membrane integrity statistics; (C) Acrosomal integrity of sperm detected by FITC-PNA staining (scale, 20 μm). a: sperm with intact acrosome, b: sperm with damaged acrosome; (D) Acrosome integrity statistics of sperm.

原文链接：DOI: 10.1016/j.cryobiol.2026.105597