基于微流控芯片模拟生理状况下精子自然优选研究

目的:在微流控芯片上模拟生理状态下的精子与宫颈粘液相互作用过程,寻找一种自然的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,基于精子与宫颈粘液相互作用在芯片通道中模拟生理状态下的精子自然优选过程,进而实现对精子的优选,同时在芯片上集成实时精子检测池,通过计算机辅助精子分析系统(CASA)实时对精子的各项参数进行测定。本研究用微流控芯片法和常规上游法同时分析30例标本,并与分选前比较。结果:分选前a+b级精子百分率为(29.78±11.24)%、正常形态精子百分率为(8.00±5.19)%、直线速度(VSL)为(18.89±4.90)μm/s、平均路径速度(VAP)为(26.84±5.13)μm/s、前向性(STR)为(70.15±7.61)%。常规上游法分选后各项参数相应为(71.65±11.18)%、(14.95±6.79)%、(24.14±5.95)μm/s、(32.61±6.36)μm/s、(73.87±9.34)%。微流控芯片分选后各项参数相应为(92.37±6.33)%、(23.33±7.67)%、(34.03±16.78)μm/s、(38.73±16.40)μm/s、(84.91±12.56)%。芯片法与分选前比较,各参数差异有统计学意义(P均<0.01);与上游法比较,各参数差异亦有统计学意义(P均<0.05)。结论:本研究在微流控芯片上模拟生理状态下精子与宫颈粘液的相互作用过程,实现了对精子的自然优选和实时检测。优选出的精子质量较分选前及上游法都有明显提高,为芯片上实现模拟生理状态下的受精过程奠定了基础。

Sperm sorting based on the imitation of the physiological process on the microfluidic chip

Objective: To establish a new method for sperm sorting by imitating the physiological process of sperm-cervical mucus interaction on the microfluidic chip.Methods: We designed a microfluidic chip to imitate the physiological process of natural sperm sorting in the microchannel based on the interaction between sperm and cervical mucus,and obtained motile sperm after the interaction.Meanwhile,we established an integrated real-time sperm detection reservoir on this chip to determine sperm parameters using the computer-assisted sperm analysis system.We analyzed 30 samples using both microfluidic and swim-up methods,and compared the results with those obtained before sorting.Results: The rate of grade a+b sperm,the rate of morphologically normal sperm,straight-line velocity(VSL),average path velocity(VAP) and straightness(STR) were(29.78±11.24)%,(8.00±5.19)%,(18.89±4.90) μm/s,(26.84±5.13) μm/s and(70.15±7.61)%,respectively,before sorting,(71.65±11.18)%,(14.95±6.79)%,(24.14±5.95) μm/s,(32.61±6.36) μm/s and(73.87±9.34)%,respectively,after swim-up sorting,and(92.37±6.33)%,(23.33±7.67)%,(34.03±16.78) μm/s,(38.73±16.40) μm/s and(84.91±12.56)%,respectively,after sorting on the microfluidic chip.The sperm parameters obtained before sorting showed statistically significant differences from those obtained on the chip(P<0.01) and by the swim-up method(P<0.05).Conclusion: Imitation of the physiological interaction between sperm and cervical mucus on the microfluidic chip helped the realization of both the natural sorting and real-time analysis of sperm.The quality of the sperm sorted on the microfluidic chip is significantly better than that of the sperm before sorting and sorted by the swim-up method.This has prepared the ground for imitating the fertilization process under the physiological condition on the microfluidic chip.