Controlled loading of cryoprotectants (CPAs) to oocyte with linear and complex CPA profiles on a microfluidic platform.
Controlled loading of cryoprotectants (CPAs) to oocyte with linear and complex CPA profiles on a microfluidic platform.
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DOI：
10.1039/c1lc20377k
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发表时间：
2011-10-21
期刊：
影响因子：
6.1
通讯作者：
Toner M
中科院分区：
文献类型：
--
作者：
Heo YS;Lee HJ;Hassell BA;Irimia D;Toth TL;Elmoazzen H;Toner M
卵母细胞冷冻保存已成为治疗不孕症的重要工具，可以为接受化疗的妇女保存卵母细胞。然而，尽管最近取得了进展，所有冷冻保存的卵母细胞的妊娠率仍然很低。在保存过程中不可避免地使用冷冻保护剂（CPA），通过CPA毒性或渗透损伤影响保存的卵母细胞的活力和妊娠率。目前的方案试图通过使CPA浓度最小化，或通过经由向细胞中逐步添加CPA使体积变化最小化来降低CPA毒性。虽然逐步添加降低了对卵母细胞的渗透压休克，但不幸的是，由于长时间暴露于CPA，它增加了毒性损伤。为了解决目前的协议的局限性，并合理地设计协议，尽量减少暴露于CPA，我们开发了一种微流控装置的定量测量卵母细胞体积在各种CPA加载协议。我们在空间上将单个卵母细胞固定在微流体装置上，创建精确控制的连续CPA曲线（逐步的、线性的和复杂的）用于向卵母细胞添加CPA，并测量卵母细胞对每个曲线的体积响应。与线性和复杂的配置文件，我们能够加载1.5 M丙二醇卵母细胞在不到15分钟，体积变化小于10%。因此，我们相信这种单卵母细胞分析技术最终将有助于辅助生殖技术和生育力保存的未来发展。
Oocyte cryopreservation has become an essential tool in the treatment of infertility by preserving oocytes for women undergoing chemotherapy. However, despite recent advances, pregnancy rates from all cryopreserved oocytes remain low. The inevitable use of the cryoprotectants (CPAs) during preservation affects the viability of the preserved oocytes and pregnancy rates either through CPA toxicity or osmotic injury. Current protocols attempt to reduce CPA toxicity by minimizing CPA concentrations, or by minimizing the volume changes via the step-wise addition of CPAs to the cells. Although the step-wise addition decreases osmotic shock to oocytes, it unfortunately increases toxic injuries due to the long exposure times to CPAs. To address limitations of current protocols and to rationally design protocols that minimize the exposure to CPAs, we developed a microfluidic device for the quantitative measurements of oocyte volume during various CPA loading protocols. We spatially secured a single oocyte on the microfluidic device, created precisely controlled continuous CPA profiles (step-wise, linear and complex) for the addition of CPAs to the oocyte and measured the oocyte volumetric response to each profile. With both linear and complex profiles, we were able to load 1.5 M propanediol to oocytes in less than 15 min and with a volumetric change of less than 10%. Thus, we believe this single oocyte analysis technology will eventually help future advances in assisted reproductive technologies and fertility preservation.
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7.4
作者：
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