Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSper...Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSperm and ISolate. Methods: This prospective study used semen specimens from 22 patients. Specimens from nine patients were processed by both PureSperm density gradient and swim-up method. These specimens were then cryopreserved. Thirteen specimens were processed by both PureSperm (40 % and 80 %) and Isolate (50 % and 90 %) double density gradient techniques. The two fractions processed by both PureSperm and swim-up were analyzed for post-wash sperm characteristics. Post-thaw analysis was done after 24 hours. Sperm fractions obtained after processing with PureSperm and ISolate were compared for post-wash sperm characteristics and ROS levels. Results: Specimens prepared with PureSperm had significantly higher median total motile sperm counts (TMSC) (32.2 x 10~6 vs. 17.6 x 10~6), recovery rates (69.2 % vs. 50.0 %), and longevity at 4 hours (83.0 % vs. 55.0 %) compared to specimen prepared by swim-up. Post-thaw specimens also had a higher recovery and longevity at 4 hours with PureSperm as compared to the swim-up. Semen specimens processed by PureSperm had significantly higher total sperm count, TMSC, and percentage recovery rates (30.0 % vs. 19.7 %) than ISolate. Conclusion: Semen quality is better preserved in fresh and cryopreserved semen prepared with PureSperm density gradient compared to swim-up. A significant enrichment of sperm is observed with PureSperm compared to ISolate. Higher recovery rates of mature motile sperm obtained after PureSperm sperm preparation may be beneficial for successful ART. i展开更多
目的:评价1层和2层Percoll密度梯度离心法分离精子的效果。方法:20份精液标本分别行50%1层,90%和45%Percoll2层密度梯度离心分离,处理前后应用SCA(sperm class analyzer)精子质量分析仪分析精子密度、活力和圆形细胞密度。结果:1层法分...目的:评价1层和2层Percoll密度梯度离心法分离精子的效果。方法:20份精液标本分别行50%1层,90%和45%Percoll2层密度梯度离心分离,处理前后应用SCA(sperm class analyzer)精子质量分析仪分析精子密度、活力和圆形细胞密度。结果:1层法分离后精子回收率为(65.5±12.8)%,明显高于2层法(P<0.01);1层和2层法分离后a级精子百分率明显高于处理前(P<0.05,P<0.01),而1层法分离后a级精子百分率明显低于2层法(P<0.05);1层法分离精子后c级精子百分率明显高于2层法(P<0.05),与处理前相比没有明显差异(P>0.05);2层法分离后a+b级精子百分率明显高于处理前(P<0.05),1层法分离后a+b级精子百分率与处理前相比没有明显差异(P>0.05);1层和2层法分离后圆形细胞密度明显低于处理前(P<0.05,P<0.01),两种方法之间没有差异(P>0.05)。结论:1层法分离后精子回收率较高,精子的活力改变不大;2层法分离后精子回收率较低,精子的活力明显改善;1层和2层法都可以较好地把精子与圆形细胞分开。两种方法各有优势,在精子体外处理中都有着重要的应用价值。展开更多
Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A tot...Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay （SCSA） parameters, that is, DNA fragmentation index （%DFI） and high DNA stainability （%HDS）, were assessed on the flesh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation （DGC） was followed by swimup treatment in comparison with DGC alone （P 〈 0.01）. Although the %DFI increased in a time-dependent manner with incubation both at room temperature （RT） and at 37℃ in air, the %DFI after 24 h at RT was significantly lower than that at 37℃ （P 〈 0.05）. Incubation with 5% CO2 was effective in maintaining sperm motility （P 〈 0.01）; however, it induced further elevation of %DFI （P 〈 0.001）. Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.展开更多
文摘Aim: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSperm and ISolate. Methods: This prospective study used semen specimens from 22 patients. Specimens from nine patients were processed by both PureSperm density gradient and swim-up method. These specimens were then cryopreserved. Thirteen specimens were processed by both PureSperm (40 % and 80 %) and Isolate (50 % and 90 %) double density gradient techniques. The two fractions processed by both PureSperm and swim-up were analyzed for post-wash sperm characteristics. Post-thaw analysis was done after 24 hours. Sperm fractions obtained after processing with PureSperm and ISolate were compared for post-wash sperm characteristics and ROS levels. Results: Specimens prepared with PureSperm had significantly higher median total motile sperm counts (TMSC) (32.2 x 10~6 vs. 17.6 x 10~6), recovery rates (69.2 % vs. 50.0 %), and longevity at 4 hours (83.0 % vs. 55.0 %) compared to specimen prepared by swim-up. Post-thaw specimens also had a higher recovery and longevity at 4 hours with PureSperm as compared to the swim-up. Semen specimens processed by PureSperm had significantly higher total sperm count, TMSC, and percentage recovery rates (30.0 % vs. 19.7 %) than ISolate. Conclusion: Semen quality is better preserved in fresh and cryopreserved semen prepared with PureSperm density gradient compared to swim-up. A significant enrichment of sperm is observed with PureSperm compared to ISolate. Higher recovery rates of mature motile sperm obtained after PureSperm sperm preparation may be beneficial for successful ART. i
文摘目的:评价1层和2层Percoll密度梯度离心法分离精子的效果。方法:20份精液标本分别行50%1层,90%和45%Percoll2层密度梯度离心分离,处理前后应用SCA(sperm class analyzer)精子质量分析仪分析精子密度、活力和圆形细胞密度。结果:1层法分离后精子回收率为(65.5±12.8)%,明显高于2层法(P<0.01);1层和2层法分离后a级精子百分率明显高于处理前(P<0.05,P<0.01),而1层法分离后a级精子百分率明显低于2层法(P<0.05);1层法分离精子后c级精子百分率明显高于2层法(P<0.05),与处理前相比没有明显差异(P>0.05);2层法分离后a+b级精子百分率明显高于处理前(P<0.05),1层法分离后a+b级精子百分率与处理前相比没有明显差异(P>0.05);1层和2层法分离后圆形细胞密度明显低于处理前(P<0.05,P<0.01),两种方法之间没有差异(P>0.05)。结论:1层法分离后精子回收率较高,精子的活力改变不大;2层法分离后精子回收率较低,精子的活力明显改善;1层和2层法都可以较好地把精子与圆形细胞分开。两种方法各有优势,在精子体外处理中都有着重要的应用价值。
文摘Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay （SCSA） parameters, that is, DNA fragmentation index （%DFI） and high DNA stainability （%HDS）, were assessed on the flesh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation （DGC） was followed by swimup treatment in comparison with DGC alone （P 〈 0.01）. Although the %DFI increased in a time-dependent manner with incubation both at room temperature （RT） and at 37℃ in air, the %DFI after 24 h at RT was significantly lower than that at 37℃ （P 〈 0.05）. Incubation with 5% CO2 was effective in maintaining sperm motility （P 〈 0.01）; however, it induced further elevation of %DFI （P 〈 0.001）. Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.
