目的 在微流控芯片上模拟精子与宫颈粘液相互作用过程,发展一种接近生理状态的精子优选方法。 方法 按功能构建微流控芯片,使精子在灌注有宫颈粘液的通道中运行。同时在芯片上集成在线精子检测池,对精子的各项参数进行在线测定。用微流控芯片法和常规上游法对照分析35例标本并比较分选效果。 结果 芯片法和上游法分选都可以显著改善精子的质量(P<0.01)。实验对两种方法分选后精子的各项参数进行了比较。与上游法相比,芯片法分选精子在精子活力(93.41±6.02% vs 70.66±10.81%)、正常形态百分率为(22.28±7.42%vs 15.05±6.57%)、平均轨迹速度(41.81±15.87μm/s vs 34.58±6.15μm/s)、前向性比例(85.02±12.18%vs 70.85±9.03%)等方面均具有显著性差异(P<0.01),而两种方法分选获得的精子在直线运动速度(35.08±16.32μm/s vs 23.18±5.75μm/s)比较中没有显著差异(P>0.05)。 结论 本研究在微流控芯片上实现了精子的自然优选。这种基于芯片的精子分选方法可以显著改善精子质量。与传统上游法相比,芯片法精子分选在多项参数上均具有优势。%Objective To develop a microfluidic chip based sperm sorting method by mimicking the interaction between sperm and cervical mucus. Methods A microfluidic chip was designed and fabricated, which included a sample reservoir, a retrieving reservoir that linked by a microchannel. Before adding raw sperm sample to the sample reservoir, the microchannel was filled with cervical mucus. Sperms were sorted by allowing them to pass through the microchannel. Sperms with high motility were obtained in the retrieving reservoir through their interaction with the mucus. The sorted sperms were further tested on the microchip using a computer-assisted sperm analysis system. Thirty-five sperm samples were processed by the microfluidic method and the conventional swimming-up method in parallel, and results obtained from the two methods were compared. Results Both the microfluidic and the swimming-up method was able to increase the percentage of high-motility sperms (P<0.01). The microfluidic method was superior to the swimming-up method in terms of sperm motility (93.41±6.02%vs 70.66±10.81%), sperm normal morphology (22.28±7.42%vs 15.05±6.57%), average path velocity (41.81±15.87 μm/s vs 34.58±6.15 μm/s) and straightness (85.02±12.18% vs 70.85±9.03%) . These two methods didn't show significant difference in straight-line velocity (35.08±16.32μm/s vs 23.18±5.75μm/s, P>0.05). Conclusion By taking advantage of the interaction between sperms and cervical mucus, physiological sperm sorting was achieved on a microfluidic chip. This microfluidic method was able to improve sperm quality and showed advantage over the conventional method.
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