基于微流控芯片的动态胚胎培养

目的:在微流控芯片上模拟输卵管收缩和纤毛运动引起的流体机械刺激和生化刺激,发展一种接近生理状态的胚胎动态培养方法,探讨其在体外受精-胚胎培养中的价值。方法:按功能构建微流控芯片,优化流体速度,使小鼠受精卵在芯片微孔中接受持续的灌注培养,同时在芯片上实现胚胎发育的监测。比较用微流控芯片法和常规微滴培养法的胚胎发育情况和囊胚形成率。结果:微流控培养系统流体速度在10μl/h时胚胎发育情况较好,获得的囊胚率最高(47.9%,23/48)。体外受精胚胎在微流控培养组5~8细胞发育率、桑葚胚发育率和囊胚形成率均明显高于微滴培养组(53.1%/46.6%、50.3%/41.5%和45.5%/35.5%),两组之间比较差异均有统计学意义(P均0.05)。而两种培养方法获得的2~4细胞发育率分别为63.4%和60.2%,差异无统计学意义。结论:胚胎培养过程中的物理微环境会影响胚胎质量,这种基于微流控芯片的胚胎动态培养方法可以显著改善胚胎质量,有希望成为辅助生殖技术中胚胎培养的一种新型工具。

Dynamic embryo culture in a microfluidic device

Objective.. To develop a microfluidic chip for embryo dynamic culture method by mimicking the fluid-mechanical and biochemical stimulation embryos experience in vivo from ciliary currents and oviductal contractions. Methods： A microfluidic chip was designed and fabricated for testing the influence of continual fluid flow on embryo development. This gave rise to design of a microfluidics system using microchannels as conduits for fluid flow through a 16 - microcellular where embryo resided. Mouse embryos were cultured in conventional microdrop-static control and dy- namic conditions with microfluidics, and results obtained from the two methods were compared. Results.. Embryonic development situation was the best and the blastocyst rate was the highest （47.9%, 23/48） when fluid velocity of microfluidic culture system was 10 u 1 per hour. The microfluidic chip dynamic culture method significantly improved the embryo development. The microfluidic dynamic method was superior to the microdrop-static method in terms of 5 to 8 cell embryo rate （53.1% vs. 46.6%）, morula rate （50.3% vs. 41.5%） and blastocyst rate （45.5% vs. 35.5%, P all 0.05）. There was no significant difference in the rate of 2 to 4 cell-embryo rate between these two methods （63.4% vs. 60.2%, P 〉0.05）. Conclusion The physical environment in the process of embryo culture can affect the quality of embryo. This microfluidic method maybe able to improve embryo development and show an advantage over the conven- tional method, which would be expected to serve as a powerful tool for embryo culture in the future.