Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes

Yang Yun; Zhou Xinli; Dai Jianjun; Zhang Defu; Yi Xingyue; Teng Yun; Tao Leren

doi:10.3969/j.issn.0253-4339.2017.03.114

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Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes

更新时间：2024-07-18

- Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes
- Journal of Refrigeration Vol. 38, Issue 3, (2017)
- 作者机构：
  
  1. 上海理工大学生物热科学研究所
  2. 上海市农业科学院畜牧兽医研究所
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.0253-4339.2017.03.114
  CLC：
- Published：2017
- 稿件说明：
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Yang Yun, Zhou Xinli, Dai Jianjun, et al. Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes[J]. Journal of refrigeration, 2017, 38(3).
DOI：

Yang Yun, Zhou Xinli, Dai Jianjun, et al. Osmotic Injury to Porcine Oocytes during Cryoprotective Agents Addition and Removal Processes[J]. Journal of refrigeration, 2017, 38(3). DOI： 10.3969/j.issn.0253-4339.2017.03.114.

摘要

在卵母细胞低温保存过程中，低温保护剂的添加与去除是必不可少的步骤，但此过程会对细胞造成致命的渗透损伤和毒性损伤。为了研究保护剂添加和去除联合过程对猪MII期细胞的损伤程度，本文设计并制作了一个适合卵母细胞低温保护剂连续添加与去除的微流体装置，分别采用微流控线性法和传统的分步法加载和去除低温保护剂，分析了两种方法对卵母细胞的体积变化以及存活率与发育潜能的影响。结果表明：微流控线性法加载和去除低温保护剂时，卵母细胞的体积变化明显小于分步法；线性法处理后细胞的存活率、卵裂率、囊胚率分别为95.3%、64.4%、19.4%，都显著高于传统的分步法（79.4%、43.6%、9.7%）（P<0.05）。因此，微流体装置用于卵母细胞低温保护剂添加与去除是有效的，能够显著减小细胞的渗透损伤，为卵母细胞低温保存技术提供了新思路。

Abstract

Oocyte cryopreservation has become a practical tool in assisted reproductive technology and fertility preservation. Cryoprotective agents (CPAs) addition and removal are essential steps during oocytes cryopreservation

however

the processes of CPAs addition and removal may cause fatal osmotic and toxic injuries to oocytes. In order to study the injuries of porcine MII-stage oocytes during CPAs addition and removal combined processes and minimize osmotic and toxic injuries to oocytes

primarily

a microfluidic device for continuous loading and unloading of CPAs was designed and fabricated in this study. CPAs were loaded and unloaded with the microfluidic linear method and the conventional step-wise method

respectively. Then

the cell volume changes and the effects on the survival rate and developmental potential of oocytes were investigated. The results showed that the oocyte volume changes with the microfluidic device were obviously less than step-wise method. 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 be used efficiently to load and unload of CPAs

and significantly reduce the osmotic shock to oocytes

which may open up a new path for oocyte cryopreservation.

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Related Institution

上海理工大学

Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology, Shanghai Technical Service Platform for Cryopreservation of Biological Resources, Shanghai Collaborative Innovation Center for Tumor Treatment with Energy

Institute of Biothermal and Technology, University of Shanghai for Science and Technology

Institute of Biothermal Technology, University of Shanghai for Science and Technology

Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology

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