Microfluidic method reduces osmotic stress injury to oocytes during cryoprotectant addition and removal processes in porcine oocytes |
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| Affiliation: | 1. IVI Valencia, IVI-RMA Global, Plaza Policía Local, 3, 46015, Valencia, Spain;2. Embryotools, Parc Científic de Barcelona, Av. Doctor Marañon, 8, 08028, Barcelona, Spain;1. Sydney School of Veterinary Science, Faculty of Science, The University of Sydney, Camden, NSW 2570, Australia;2. Westmead Fertility Centre, Westmead Hospital, Westmead, NSW 2145, Australia;1. Graduate School of Science and Technology, Shinshu University, Ueda, Nagano, 386-8567, Japan;2. Graduate School of Medicine, Science and Technology, Shinshu University, Ueda, Nagano, 386-8567, Japan;3. Faculty of Textile Science and Technology, Shinshu University, Nagano, 386-8567, Japan;4. National Institute for Physiological Sciences, Okazaki, Aichi, 444-8787, Japan;5. School of Life Science, The Graduate University for Advanced Studies, Okazaki, Aichi, 444-8787, Japan;1. Department of Mechanical Engineering, Villanova University, Villanova, PA 19085, USA;2. Institute of Molecular Medicine and Genetics, Department of Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA 30912, USA;3. School of Chemical, Biological and Environmental Engineering, Oregon State University, Corvallis, OR 97331, USA;4. Department of Obstetrics and Gynecology, and Cancer Center, Medical College of Georgia, Georgia Regents University, Augusta, GA 30912, USA;1. Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Yunnan Animal Science and Veterinary Institute, Kunming, Yunnan 650224, People''s Republic of China;2. College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan 650201, People''s Republic of China |
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| Abstract: | 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. |
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| Keywords: | Oocyte Microfluidics CPA Osmotic damage Cryopreservation |
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