| 摘要: |
| 卵母细胞玻璃化保存的需求日益增加,目前人工或自动化保存装置分步加载和去除低温保护剂易造成卵母细胞的渗透 损伤和丢失。 本研究采用低成本易制作的 PMMA 微流控芯片,结合石英毛细管搭建微流控混合系统,既实现了保护剂的连续加 载/ 去除,又为后续浸入液氮实现自动玻璃化保存做准备。 通过采用手动多步法和微流控法对小鼠 MⅡ期卵母细胞加载/ 去除低 温保护剂,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。 |
| 关键词: 玻璃化保存 微流控 卵母细胞 石英毛细管 PMMA 芯片 |
| DOI： |
| 投稿时间：2022-01-20 修订日期：2022-03-26
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| 基金项目:本文受上海市促进市级医院临床技能与临床创新能力三 年行动计划重大临床研究项目( SHDC2020CR3077B) 资助。 |
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| Microfluidic System for Vitrification of Oocytes |
| Zhang Yuqi,Yu Zixuan,Zhou Xinli |
| (Institute of Biothermal Technology, University of Shanghai for Science and Technology) |
| 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. |
| Key words: vitrification microfluidics oocyte quartz capillary PMMA chip |
