Genotoxicity assessment of mouse oocytes by comet assay before vitrification and after warming with three vitrification protocols

被引:17
作者
Berthelot-Ricou, Anais [1 ,2 ]
Perrin, Jeanne [1 ,3 ]
di Giorgio, Carole [5 ]
de Meo, Michel [5 ]
Botta, Alain [1 ]
Courbiere, Blandine [1 ,4 ]
机构
[1] Aix Marseille Univ, IMBE, ECCOREV, Santee Humaine & Environm UMR 6116,FR CNRS 309, Marseille, France
[2] CHU La Reunion, Dept Gynecol, CH Felix Guyon, St Denis, Reunion, France
[3] AP HM La Concept, Dept Gynecol Obstet & Reprod, CECOS Reprod Lab, F-13005 Marseille, France
[4] AP HM La Concept, Dept Gynecol Obstet & Reprod, 147 Bd Baille, F-13005 Marseille, France
[5] Fac Pharm Marseille, Lab Environm Mutagenesis, IMBE, Marseille, France
关键词
Oocyte vitrification; DNA damage; cryoprotectants; comet assay; genotoxicity; IN-VITRO; CRYOPRESERVATION; 1,2-PROPANEDIOL; CONFIGURATION; FERTILITY; INCREASES; SURVIVAL; SPINDLE; RATES;
D O I
10.1016/j.fertnstert.2013.05.025
中图分类号
R71 [妇产科学];
学科分类号
100211 ;
摘要
Objective: To assess the genotoxicity of three oocyte vitrification protocols. Design: Murine assay. Setting: Biogenotoxicology research laboratory. Animal(s): CD1 female mice. Intervention(s): Three mouse oocyte groups were exposed to three commercialized human oocyte vitrification protocols. Protocols 1 and 2 contained dimethyl sulfoxide and ethylene glycol (EG), and protocol 3 contained EG and 1,2-propanediol (PrOH). DNA damage was first evaluated by comet assay after oocyte exposure to the three different equilibration and vitrification solutions. Comet assay was also performed after full vitrification and warming procedure and compared with a negative control group (oocytes stored in medium culture only) and a positive control group (oocytes exposed to hydrogen peroxide just before comet assay). Main Outcome Measure(s): DNA damage was quantified as Olive tail moment (OTM). Statistical analysis consisted of a Shapiro-Wilk test. Then, median protocol OTM was compared with the negative control group with the Mann-Whitney U test. The difference was considered to be statistically significant if the P value was <.05. Result(s): In both parts of our study, protocols 1 and 2 did not induce significant DNA damage, whereas protocol 3 induced statistically higher DNA damage compared with the negative control group. Conclusion(s): Vitrification protocols containing PrOH induced significant DNA damage on mouse oocytes, both before cooling and after warming. Therefore, for the moment, we prefer vitrification techniques without PrOH while we await more studies on PrOH toxicity and long-term evaluation. (C) 2013 by American Society for Reproductive Medicine.
引用
收藏
页码:882 / 888
页数:7
相关论文
共 35 条
[21]   Calcium-free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes [J].
Larman, MG ;
Sheehan, CB ;
Gardner, DK .
REPRODUCTION, 2006, 131 (01) :53-61
[22]   Successful ongoing pregnancies after vitrification of oocytes [J].
Lucena, E ;
Bernal, DP ;
Lucena, C ;
Rojas, A ;
Moran, A ;
Lucena, A .
FERTILITY AND STERILITY, 2006, 85 (01) :108-111
[23]   Practical issues on the application of the GHS classification criteria for germ cell mutagens [J].
Morita, Takeshi ;
Hayashi, Makoto ;
Nakajima, Madoka ;
Tanaka, Noriho ;
Tweats, David J. ;
Morikawa, Kaoru ;
Sofuni, Toshio .
REGULATORY TOXICOLOGY AND PHARMACOLOGY, 2009, 55 (01) :52-68
[24]   Over 900 oocyte cryopreservation babies born with no apparent increase in congenital anomalies [J].
Noyes, N. ;
Porcu, E. ;
Borini, A. .
REPRODUCTIVE BIOMEDICINE ONLINE, 2009, 18 (06) :769-776
[25]   Oocytes or embryo storage? [J].
Porcu, E ;
Fabbri, R ;
Ciotti, PM ;
Frau, F ;
De Cesare, R ;
Venturoli, S .
FERTILITY AND STERILITY, 2002, 78 (03) :S15-S15
[26]   Evaluating methods of mouse euthanasia on the oocyte quality: cervical dislocation versus isoflurane inhalation [J].
Roustan, Audrey ;
Perrin, Jeanne ;
Berthelot-Ricou, Anais ;
Lopez, Erica ;
Botta, Alain ;
Courbiere, Blandine .
LABORATORY ANIMALS, 2012, 46 (02) :167-169
[27]   Cryopreservation of supernumerary oocytes in IVF/ICSI cycles [J].
Setti, PEL ;
Albani, E ;
Novara, PV ;
Cesana, A ;
Morreale, G .
HUMAN REPRODUCTION, 2006, 21 (02) :370-375
[28]   Morphological changes, DNA damage and developmental competence of in vitro matured, vitrified-thawed buffalo (Bubalus bubalis) oocytes: A comparative study of two cryoprotectants and two cryodevices [J].
Sharma, G. Taru ;
Dubey, Pawan K. ;
Chandra, Vikash .
CRYOBIOLOGY, 2010, 60 (03) :315-321
[29]   Fundamental cryobiology of mammalian oocytes and ovarian tissue [J].
Shaw, JM ;
Oranratnachai, A ;
Trounson, AO .
THERIOGENOLOGY, 2000, 53 (01) :59-72
[30]   The comet assay as an indicator test for germ cell genotoxicity [J].
Speit, Guenter ;
Vasquez, Marie ;
Hartmann, Andreas .
MUTATION RESEARCH-REVIEWS IN MUTATION RESEARCH, 2009, 681 (01) :3-12