title: High survival of mouse oocytes using an optimized vitrification protocol.
abstract: The method of vitrification has been widely used for cryopreservation. However, the effectiveness of this method for mammalian oocytes could be improved by optimizing each step of the process. In the present study, we tested the effects of varying several key parameters to determine the most effective protocol for mouse oocyte vitrification. We found that cryoprotectant containing ethylene glycol and dimethylsulfoxide plus 20% fetal calf serum produced the highest rates of oocyte survival, fertilization, and blastocyst formation. The duration and temperature of oocyte exposure to vitrification and thawing solutions influenced survival rate. The presence of cumulus cells surrounding oocytes and the incubation of thawed oocytes in Toyoda-Yokoyama-Hosoki medium also increased oocyte survival. Open pulled straw and nylon loop methods were more effective than the mini-drop method. Finally, the combination of these improved methods resulted in better spindle morphology when compared to the unimproved methods. These results demonstrate that the outcomes of mouse oocyte vitrification can be improved by a suitable combination of cryopreservation methods, which could be applied to future clinical research with human oocytes.
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Cheng-Jie
authors.initials: CJ
authors.lastname: Zhou
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Dong-Hui
authors.initials: DH
authors.lastname: Wang
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Xin-Xin
authors.initials: XX
authors.lastname: Niu
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Xiang-Wei
authors.initials: XW
authors.lastname: Kong
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Yan-Jiao
authors.initials: YJ
authors.lastname: Li
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Ren
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Zhou
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Angeleem
authors.initials: A
authors.lastname: Lu
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Yue-Fang
authors.initials: YF
authors.lastname: Zhao
authors.affiliation: The Key Laboratory of National Education, Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.
authors.forename: Cheng-Guang
authors.initials: CG
authors.lastname: Liang
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005306:Fertilization / Q000187:drug effects / Q000502:physiology; D005307:Fertilization in Vitro / Q000187:drug effects / Q000379:methods; D008297:Male; D051379:Mice; D009865:Oocytes / Q000166:cytology*; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide