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| Experimental Study of Linear Removing Cryoprotective Agents from MII Porcine Oocytes with Microfluidics Device |
| Zhou Xinli1*, Yang Yun1, Dai Jianjun2, Zhang Defu2, Shao Wenqi1, Yi Xingyue1, Tao Leren1 |
1Institute of Biothermal Technology, University of Shanghai for Science and Technology, Shanghai 200093, China;
2Animal and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China |
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Abstract Cryoprotective agents (CPAs) must be removed from cryopreserved oocytes before used in clinic. Step-wise methods usually lead to the cell loss due to complicated operating steps, and may cause fatal osmotic injury to oocytes during CPAs removal process. In order to minimize the osmotic injury to oocytes, a microfluidic device for unloading CPAs from oocytes was designed and fabricated in this study. CPAs were linear unloaded from MII porcine oocytes with microfluidic device under different durations (6 min, 8 min and 10 min), the cell volume changes and the effects on the survival and developmental rate of oocytes were investigated, and then compared with that obtained by the conventional step-wise methods. Results showed that 8 min duration was optimal for linear unloading CPAs with microfluidic device. The linear method remarkably reduced the osmotic injury to oocytes during the removal of CPAs. The highest normalized swelling volume of oocyte only reached 1.12±0.07. The survival, cleavage and blastocyst rate of oocytes were 83.6%, 72.4% and 21.5%,respectively, which were significantly higher than those of one-step method and step-wise methods (P<0.05). In conclusion, linear unloading CPAs with the microfluidic method can significantly alleviate the osmotic damage to oocytes, which may provide a new path for oocyte cryopreservation.
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Received: 20 September 2016
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