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目的:建立一种接近生理状态的胚胎动态培养方法。方法:按功能设计和构建微流控芯片,使小鼠体外受精卵在微流控芯片微孔中接受持续的灌注培养,同时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,并与常规微滴培养法比较,监测胚胎发育情况和囊胚形成率。结果:微流控芯片动态培养方法可显著改善胚胎的质量。与常规微滴培养法相比,微流控动态培养获得的4-细胞胚胎、桑椹胚和囊胚形成率均有显著提高(68.4±1.2%vs 53.2±2.5%;55.3±2.6%vs 45.5±3.3%;40.5±2.7%vs 35.5±2.3%)(P<0.05)。而2种方法获得的卵裂率无统计学差异(75.5±3.2%vs 73.9±4.2%,P>0.05)。结论:在微流控芯片上实现了小鼠近生理状态的胚胎培养,显著改善了胚胎质量,有希望成为一种胚胎培养的有力工具。
Objective: To establish a method of dynamic culture of embryos close to physiological state. Methods: The microfluidic chip was designed and constructed according to the function, the mouse in vitro fertilized egg was subjected to continuous perfusion culture in the microfluidic micro-hole, Simultaneous simulation of tubal contraction and ciliary motility caused by fluid mechanical stimulation and biochemical stimulation, and Conventional microdialysis methods were compared to monitor embryonic development and blastocyst formation rates. Results: Microfluidic chip dynamic culture method can significantly improve the quality of embryos. Compared with the conventional droplet culture method, the formation rates of 4-cell embryos, morula and blastocysts obtained by microfluidic dynamic culture were significantly increased (68.4 ± 1.2% vs 53.2 ± 2.5%; 55.3 ± 2.6% vs 45.5 ± 3.3 %; 40.5 ± 2.7% vs 35.5 ± 2.3%) (P <0.05). The cleavage rates obtained by the two methods were not statistically different (75.5 ± 3.2% vs 73.9 ± 4.2%, P> 0.05). Conclusion: The mouse embryo culture in the physiological state is realized on the microfluidic chip, the quality of the embryo is remarkably improved, and it is hopeful to be a powerful tool for embryo culture.