Investigation of experiment in improving of rapid cryopreservation carrier on scarce human spermatozoa
Chen Guoyong LiWen
1.Reproductive Medicine Center , Changzheng Hospital, Naval Medical University of the PLA, Shanghai 200003, China; 2. Reproductive Medicine Center, 900 Hospital of the Joint Logistics Team , Fujian Fuzhou, 350025,China
Abstract: 【Abstract】 Objective To explore the investigation of experiment in improving of rapid cryopreservation carrier on scarce human spermatozoa, and provide reference for cryopreservation of sperm in patients withsevere oligo-asthenozoospermia. Methods From December 2018 to August 2019, male infertility treated in center of Reproductive Medicine, 900 Hospital of the Joint Logistics Team were divided into the normal sperm group and oligo-asthenospermic group through their semen quality. Three samples of sperm were selectedfrom each experimental sample and were mixed and diluted toconcentration (1-2)×106/ml, 27 mixed cases were selected from 81 samples of normal semen, 18 mixed cases were selected from 54 oligoasthenic sperm samples. According to different frozen carrier spermatozoon were frozen in 3 groups: thin slice sperm tube (group A), wheat tube micro-volume freezing group (group B), modified rapid freezing carrier group(group C). All of them used commercialized sperm freezing reagent 1 ∶ 1 to mix, rapid cryopreservation afterfumigation. Survival rate, DNA fragmentation index, acrosome enzyme activity and normal morphology rate after cryopreservation of three scarce sperm cryopreservation carrier were compared to evaluate the freezing efficiency and safety of rapid cryopreservation carrier. Statistical methods were carried out by t-test, anova analysis and LSD multiple comparison. Results Sperm quality was decreased in normal sperm group, three cryopreservation methods were used. Group C were higher than that in group A and group B in the survival rate of resuscitation sperm motility for the normal sperm group with statistical significance (P<0.05). Group C was slightly lower than that in group A and group B in DNA fragment index of resuscitated sperm for the normal sperm group with no statistical significance (P>0.05). Normal morphologic rate after cryopreservation in the group A, B and C was lower than that before cryopreserved with statistical significance (P<0.05). Oligo-asthenospermic group: DNA fragment index of resuscitated sperm in both groups A and B was higher than that before cryopreservation with statistical significance (P<0.01). DNA fragment index of resuscitated sperm in group C was higher than that before cryopreservation but with no statistical significance (P=0.068). DNA fragment index of resuscitated sperm in group C was slightly lower than that in group A and group B with no statistical significance (P>0.01). The normal morphology rate of frozen sperm in group A, B and C was lower than that before freezing with no statistical significance (P>0.05). Normal sperm group: the acrosomal enzyme activity of sperm was (123.6±12.8) IU/106 sperm before freezing, and decreased in group A (74.7±15.6), B (84.7±13.5) and C (91.2±16.2) IU/106 after resuscitation with statistical significance (F=37.896，P<0.001). Group B and C were superior to group A in acrosomal influence of sperm withstatistical significance(P=0.043, P=0.001). Conclusion The improved rapid cryopreservation carrier has certain advantages for scarce human spermatozoa, which is worthy of further study.
2020, Vol. 8 
