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本文研究了冷冻及复温过程中人精子超微结构的变化.用甘油-卵黄-三羟甲基氨基甲烷-葡萄糖-柠檬酸混合液或纯甘油作精子冷冻保护剂,采用阶段降温法。标本在-196℃液氮中贮存一周后.37℃水浴中复温5～10分钟.透射电镜(TEM)下观察.精子超微结构变化主要在头部,可见顶体前段及质膜肿胀,顶体外膜破裂,顶体物质流失;甚至顶体内膜及核膜破损,丧失成为裸核精子。与新鲜精液相比.有完整质膜.顶体的精子百分比明显下降(P<0.01)。线粒体基质局部透亮,呈空泡状改变,并可见细小颗粒物质沉积.尾部轴丝“9+2”结构存在;质膜和核之间可见尾结构。精子头部结构完整率与精子存活率呈正相关(r=0.88 P<0.01).结果表明同一供者的冻精作人工授精,其成功率必然低于鲜精的成功率;它也可能是一般冻精人工授精成功率低于鲜精的原因.
In this paper, the ultrastructural changes of human sperm during freezing and rewarming were studied.The phase cooling method was adopted by using glycerol-yolk-trimethylamine-glucose-citric acid mixture or pure glycerol as sperm cryoprotectant. Specimens were stored in liquid nitrogen at -196 ℃ for one week, rewarmed in water bath at 37 ℃ for 5-10 minutes, observed by transmission electron microscopy (TEM), ultrastructural changes of the sperm mainly in the head, visible anterior part of the acrosome and plasma membrane swelling, Acrosome membrane rupture, loss of acrosome; even acrosomal and nuclear membrane damage, loss of become naked nuclear sperm. Compared with fresh semen, there was an intact plasma membrane, and the acrosome sperm percentage was significantly decreased (P <0.01). Mitochondrial matrix translucent, vacuole-like changes, and the deposition of small particulate matter can be seen.The “9 + 2” structure of the axial axons exists; the tail structure can be seen between the plasma membrane and the nucleus. The sperm head structure integrity rate was positively correlated with sperm survival (r = 0.88 P <0.01) .The results showed that the same donor of frozen sperm for artificial insemination, the success rate is bound to be lower than the success rate of fresh sperm; it may also be generally frozen Fine artificial insemination success rate is lower than the fresh fine reason.