微流控芯片在人类精子优选中的应用

Application of microfluidic chip in the selection of human sperm

目的研究自制微流控芯片优选人类精子的效果。方法自行构建微流控芯片装置。收集男性不育患者的精液样本:(1)分别置于无毒灭菌离心管与微流控芯片中1h、2h、3h、4h,比较各时间点两组精子活力的变化;(2)分别用微流控芯片优选精子40min、60min、80min,比较各时间点优选前后各项精子参数的变化;(3)分别用微流控芯片、上游法和密度梯度离心法优选精子,比较三种方法优选前后各项精子参数的变化。结果(1)静置1h、2h、3h、4h后,无毒灭菌离心管与微流控芯片中的精子活动率与前向运动精子百分率均无显著性差异(P>0.05)。(2)与优选前相比,微流控芯片优选精子40min、60min、80min后,精子活动率(36.54%vs 96.55%、97.92%、88.12%)和前向运动精子百分率(28.01%vs 83.04%、83.01%、81.25%)均显著上升(P<0.01),而精子浓度(47.11×10^(6)/m L vs 1.68×10^(6)/mL、3.81×10^(6)/mL、4.02×10^(6)/m L)均显著降低(P<0.01),但60min与80min组的精子浓度显著高于40min组(P<0.05);(3)与上游法和密度梯度离心法相比,微流控芯片优选后的精子活动率(89.58%vs 70.76%、71.89%)和前向运动精子百分率(82.35%vs 51.52%、45.99%)显著提升,而精子浓度(4.09×10^(6)/mL vs 17.06×10^(6)/mL、60.33×10^(6)/m L)和DFI(15.53%vs 30.08%、40.52%)显著降低(P<0.05)。结论微流控芯片对精子无毒并能优选出高质量精子,与两种常规精子优选方法相比,其优选后的精子DNA损伤较小,更适合于辅助生殖技术IVF/ICSI中使用。

Objective To evaluate the selective efficiency of human sperm using a homemade microfluidic chip.Methods Semen samples obtained from male infertile patients were respectively placed in the non-toxic sterilized centrifuge tube and microfluidic chip for 1 h,2 h,3 h,4 h and then the changes in sperm motility of the two groups were compared at each time point.Sperm samples placed in the microfluidic chip were sorted for 40 min,60 min,and 80 min,respectively and the changes of their parameters were analyzed at each time point.Sperm samples were sorted by the microfluidic chip,swim-up and density gradient centrifugation,respectively,and the changes of sperm parameters were comparatively analyzed before and after sorting.Results There were no significant differences in sperm motility and percentage of progressive sperm between the non-toxic sterilized centrifuge tube and the microfluidic chip after placing for 1 h,2 h,3 h and 4 h(P>0.05).Compared with pre-sorting,sperm motility and percentage of progressive sperm were increased significantly after sorting for 40 min,60 min and 80 min(36.54%vs 96.55%,97.92%and 88.12%;28.01%vs 83.04%,83.01%and 81.25%,P<0.01),and sperm concentration were decreased significantly(47.11×10^(6)/m L vs 1.68×10^(6)/mL,3.81×10^(6)/mL and 4.02×10^(6)/m L,P<0.01).However,sperm concentrations at 60 min and 80 min were obviously higher than that at 40 min(P<0.05).Compared with the swim-up and density gradient centrifugation,sperm motility(89.58%vs 70.76%,71.89%)and percentage of progressive sperm(82.35%vs51.52%,45.99%)sorted by the microfluidic chip were remarkably elevated,while sperm concentration(4.09×10^(6)/m L vs17.06×10^(6)/m L,60.33×10^(6)/m L)and DFI(15.53%vs 30.08%,40.52%)were significantly decreased(P<0.05).Conclusion Microfluidic chip used in the study is non-toxic to sperm and is more suitable for application in IVF/ICSI due to obtaining high-quality sperm and less DNA damage.