Institute of Biomedical Technology, University of Shanghai for Science and Technology, Shanghai, China.
Institute of Biomedical Technology, University of Shanghai for Science and Technology, Shanghai, China.
Cryobiology. 2019 Oct;90:63-70. doi: 10.1016/j.cryobiol.2019.08.005. Epub 2019 Aug 23.
Oocyte cryopreservation is an important technology in assisted reproduction and fertility preservation. However, the developmental potential of cryopreserved oocyte remains poor. Osmotic stress injury (OSI) during cryoprotectants (CPAs) loading and unloading steps has critical impact on successful cryopreservation. In order to minimize OSI to oocytes, a microfluidic device was designed and fabricated to achieve continuous CPA concentration change. MII porcine oocytes were loaded and unloaded CPAs with step-wise and microfluidic methods, oocyte volume changes were recorded and compared, loading and unloading duration of microfluidic methods were optimized. The survival and developmental rate of treated oocytes in step-wise and microfluidic linear methods were also evaluated. The results showed that oocyte volume changes with microfluidic method were obviously less than step-wise method, and the survival, cleavage and blastocyst rate of oocytes were 95.3%, 64.4%, and 19.4%, respectively, which were significantly higher than the traditional step-wise method (79.4%, 43.6%, and 9.7%) (p < 0.05). In conclusion, microfluidic device can effectively reduce the osmotic damage to oocytes and improve the survival rate and developmental rate of oocytes, which may provide a new path for oocyte cryopreservation.
卵母细胞冷冻保存是辅助生殖和生育力保存的一项重要技术。然而，冷冻保存卵母细胞的发育潜力仍然较差。在冷冻保护剂（CPAs）加载和卸载步骤中渗透胁迫损伤（OSI）对成功的冷冻保存有至关重要的影响。为了将 OSI 对卵母细胞的影响降到最低，设计并制造了一种微流控装置以实现连续的 CPA 浓度变化。使用逐步法和微流控法加载和卸载 MII 期猪卵母细胞，记录和比较卵母细胞体积变化，优化微流控法的加载和卸载时间。还评估了逐步法和微流控线性法处理卵母细胞的存活率和发育率。结果表明，微流控法的卵母细胞体积变化明显小于逐步法，卵母细胞的存活率、卵裂率和囊胚率分别为 95.3%、64.4%和 19.4%，明显高于传统的逐步法（79.4%、43.6%和 9.7%）（p < 0.05）。总之，微流控装置可以有效减少卵母细胞的渗透损伤，提高卵母细胞的存活率和发育率，为卵母细胞冷冻保存提供了新的途径。
