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The effect of two cryopreservation methods on human sperm DNA damage |
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| Institution: | 1. School of Public Health, University of Ningxia Medical, Yinchuan, 750004, People''s Republic of China;2. Institute of Toxicology, College of Preventive Medicine, University of Third Military Medical, Chongqing, 400038, People''s Republic of China;1. Department of Clinical Embryology, Central Research Lab, Kasturba Medical College, Manipal University, Manipal, 576104, Karnataka, India;2. Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, 576104, Karnataka, India;3. Department of Agriculture and Environmental Sciences, National Institute of Food Technology Enterpreneurship and Management, Kundli, India;4. Core Technology Platform, New York University, Abu Dhabi, United Arab Emirates;1. University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zatisi 728/II, 38925 Vodnany, Czech Republic;2. Sino-Czech Joint Laboratory for Fish Conservation and Biotechnology, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, China;3. University of South Bohemia in Ceske Budejovice, Faculty of Science, Institute of Chemistry and Biochemistry, Branisovska 1760, 37005 Ceske Budejovice, Czech Republic;4. Biology Centre of Academy of Sciences of the Czech Republic, Institute of Parasitology, Branisovska 31, 37005 Ceske Budejovice, Czech Republic;5. CAS Key Laboratory of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao 266101, China;1. Medical Officer, INHS Dhanvantari, C/O Navy Office, Minnie Bay, Port Blair 744102, India;2. Director and Commandant, Armed Forces Medical College, Pune 411040, India;3. Senior Advisor & Head of Department, Assisted Reproductive Technologies Centre, Army Hospital (R & R), Delhi Cantt, India |
| Abstract: | Several methods are currently available for selection when conducting sperm cryopreservation, however, these methods might cause different degrees of damage on sperm DNA. The aim of the this study is to compare the effects of storage at ?80 °C (in ultra-low temperature refrigerator) and at ?196 °C (in liquid nitrogen) on sperm DNA damage, thus to provide a reference for choosing the right method according to different aims. We randomly collected 28 semen samples from college students of Chongqing city. The samples stored at ?80 °C were neat semen samples and the samples stored at ?196 °C were mixed with additional cryoprotectants. Each sample was subjected to two freezing-thawing cycles, and the sperm DNA damage levels of fresh and thawed samples were measured by single cell gel electrophoresis (SCGE) and sperm chromatin structure assay (SCSA). Both SCGE and SCSA assays showed cryopreservation induced significant damage to sperm DNA. However, storage at ?196 °C lead to more severe damage to sperm DNA than storage at ?80 °C measured by SCSA. Sperm DNA damage increased simultaneously with the higher frequency of freezing-thawing cycles. We concluded that storage of neat semen samples at ?80 °C had milder damage to sperm DNA than storage at ?196 °C mixed with cryoprotectants. To avoid additional sperm DNA damage, repeated freezing and thawing should be prevented. |
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