ZHANG YUQI, YU ZIXUAN, ZHOU XINLI. Microfluidic System for Vitrification of Oocytes. [J]. Journal of refrigeration, 2022, 43(6). DOI: 10.3969/j.issn.0253-4339.2022.06.160.
对比卵母细胞的体积响应、存活率和后续发育率。 结果表明:微流控法注射时间 8 min 组的卵母细胞存活率、卵裂率 和囊胚率
分别达到 93. 25%、77. 12%、53. 00%
显著高于手动多步法
与对照组无显著性差异(P<0. 05)。 使用 PMMA 芯片结合 石英毛细管的微流控系统
显著减小了卵母细胞的渗透损伤和细胞丢失率
简化了操作步骤
为基于微流控技术的卵母细胞与胚 胎的自动玻璃化保存装置的开发提供关键技术支撑。
Abstract
The demand for oocyte vitrification has increased. However
the current step-by-step loading and removal of cryoprotectants by manual or automatic storage devices may cause osmotic injury and the loss of cells. In this study
low-cost polymethyl methacrylate (PMMA) microfluidic chips and quartz capillaries were used to build a microfluidic system that achieved not only the continuous loading and removal of cryoprotectants but also the subsequent immersion in liquid nitrogen to realize automatic vitrification. MII mouse oocytes were loaded or unloaded with cryoprotectants by manual multistep and microfluidic methods
and the cell volume changes
survival
and oocyte development rates were compared. The results demonstrate that the survival
cleavage
and blastocyst rates of oocytes were 93. 25%
77. 12%
and 53. 00%
respectively
for the 8 min loading / unloading duration group. These are significantly higher than those of the manual multistep group but not significantly different from those of the control group (P <0. 05). In conclusion
the microfluidic system using the PMMA chip and quartz capillaries significantly reduced osmotic injury and cell loss
and simplified the operational steps. This system may provide key technical support for the development of automated devices for oocyte and embryo vitrification based on microfluidic technology.