title: Vitrification preserves chromatin integrity, bioenergy potential and oxidative parameters in mouse embryos.
abstract: BACKGROUND
  The aim of this study was to evaluate the effects of vitrification on morpho-functional parameters (blastomere/chromatin integrity and bioenergy/oxidative potential) of mouse preimplantation embryos.
  
  
  METHODS
  In vivo produced mouse (4/16-cell, morulae and blastocyst-stage) embryos were randomly divided into vitrification and control groups. For vitrification, embryos were exposed to a 2-step loading of ethylene glycol and propylene glycol, before being placed in a small nylon loop and submerged into liquid nitrogen. After warming, the cryoprotectants were diluted by a 3-step procedure. Embryo morphology, chromatin integrity and energy/oxidative status were compared between groups.
  
  
  RESULTS
  Vitrification induced low grade blastomere cytofragmentation (P < 0.05) and low chromatin damage only in embryos at the morula stage (P < 0.001). Mitochondrial (mt) distribution pattern was affected by vitrification only in early embryos (P < 0.001). Mitochondrial activity did not change upon vitrification in morula-stage embryos but it was reduced in blastocyst-stage embryos (P < 0.05). Intracellular ROS levels significantly increased in embryos at the morula and blastocyst stages (P < 0.001). Colocalization of active mitochondria and ROS increased only in vitrified blastocysts.
  
  
  CONCLUSIONS
  In conclusion, this study elucidates the developmentally-related and mild effects of vitrification on morphology, nuclear and bioenergy/oxidative parameters of mouse embryos and demonstrates that vitrification is a suitable method for preserving predictive parameters of embryo ability to induce a full-term pregnancy.
authors.affiliation: Veterinary Clinics and Animal Productions Unit, Department of Emergency and Organ Trasplantation (DETO), University of Bari Aldo Moro, Valenzano, Bari, Italy. nicmartino@libero.it
authors.forename: Nicola A
authors.initials: NA
authors.lastname: Martino
authors.forename: Maria E
authors.initials: ME
authors.lastname: Dell'aquila
authors.forename: Rosa A
authors.initials: RA
authors.lastname: Cardone
authors.forename: Bence
authors.initials: B
authors.lastname: Somoskoi
authors.forename: Giovanni M
authors.initials: GM
authors.lastname: Lacalandra
authors.forename: Sandor
authors.initials: S
authors.lastname: Cseh
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D004734:Energy Metabolism; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D008297:Male; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D009028:Morula / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010084:Oxidation-Reduction; D011247:Pregnancy; D019946:Propylene Glycol / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D015203:Reproducibility of Results; D058989:Vitrification
chemical_list: D002843:Chromatin; D003451:Cryoprotective Agents; D017382:Reactive Oxygen Species; D019946:Propylene Glycol; D019855:Ethylene Glycol
references.citation: Fertil Steril. 2011 Jul;96(1):187-92
references.pmid: 21550043
references.citation: Nature. 2000 Nov 9;408(6809):239-47
references.pmid: 11089981
references.citation: Reprod Biomed Online. 2003 Dec;7(6):664-70
references.pmid: 14748965
references.citation: J Cell Physiol. 2010 Feb;222(2):444-55
references.pmid: 19918794
references.citation: Mol Reprod Dev. 2002 Apr;61(4):466-76
references.pmid: 11891918
references.citation: Nature. 1985 Feb 14-20;313(6003):573-5
references.pmid: 3969158
references.citation: Neurochem Res. 2008 Dec;33(12):2565-74
references.pmid: 18473170
references.citation: Birth Defects Res C Embryo Today. 2010 Sep;90(3):163-75
references.pmid: 20860056
references.citation: Trends Biochem Sci. 2005 Aug;30(8):453-61
references.pmid: 15996871
references.citation: Reprod Biomed Online. 2006 Aug;13(2):246-54
references.pmid: 16895640
references.citation: Mitochondrion. 2011 Sep;11(5):797-813
references.pmid: 20933103
references.citation: Hum Reprod. 2000 Dec;15(12):2621-33
references.pmid: 11098036
references.citation: Anim Reprod Sci. 2000 Jul 2;60-61:357-64
references.pmid: 10844207
references.citation: Fertil Steril. 2012 Mar;97(3):720-8.e1
references.pmid: 22260855
references.citation: J Neurosci. 2010 Jan 20;30(3):1015-26
references.pmid: 20089910
references.citation: Mol Reprod Dev. 2009 Nov;76(11):1056-63
references.pmid: 19551710
references.citation: Anim Reprod Sci. 2010 Jan;117(1-2):166-77
references.pmid: 19372012
references.citation: BMC Biotechnol. 2011 Mar 30;11:29
references.pmid: 21450077
references.citation: Theriogenology. 2009 Apr 15;71(7):1093-104
references.pmid: 19167745
references.citation: Reprod Biomed Online. 2004 Sep;9(3):338-47
references.pmid: 15353087
references.citation: Hum Reprod. 2002 Feb;17(2):393-406
references.pmid: 11821285
references.citation: Prog Histochem Cytochem. 2009;44(3):125-72
references.pmid: 19822255
references.citation: Free Radic Biol Med. 2000 Aug;29(3-4):222-30
references.pmid: 11035250
references.citation: Cryobiology. 2007 Oct;55(2):115-26
references.pmid: 17681290
references.citation: Mol Hum Reprod. 1996 Feb;2(2):93-8
references.pmid: 9238664
references.citation: Gynecol Endocrinol. 2010 Aug;26(8):563-7
references.pmid: 20230330
references.citation: Am J Physiol Cell Physiol. 2004 Oct;287(4):C817-33
references.pmid: 15355853
references.citation: Zygote. 2011 May;19(2):137-45
references.pmid: 20587135
references.citation: Anim Reprod Sci. 2007 Jul;100(1-2):128-40
references.pmid: 16895747
references.citation: Biol Reprod. 2002 Dec;67(6):1671-80
references.pmid: 12444040
references.citation: Biol Reprod. 2005 Mar;72(3):612-8
references.pmid: 15537860
references.citation: Hum Reprod. 1998 Apr;13(4):998-1002
references.pmid: 9619561
references.citation: Cryobiology. 1987 Oct;24(5):387-402
references.pmid: 3652721
references.citation: Acta Vet Hung. 2009 Sep;57(3):399-410
references.pmid: 19635712
references.citation: J Clin Invest. 2010 Aug;120(8):2817-28
references.pmid: 20644252
references.citation: Mol Reprod Dev. 2000 Mar;55(3):282-8
references.pmid: 10657047
references.citation: PLoS One. 2011;6(11):e27452
references.pmid: 22076161
references.citation: Am J Physiol Cell Physiol. 2010 May;298(5):C1235-44
references.pmid: 20107036
references.citation: Semin Cell Dev Biol. 2009 May;20(3):354-64
references.pmid: 19136067
references.citation: J Assist Reprod Genet. 2009 May;26(5):285-90
references.pmid: 19603263
references.citation: J Histochem Cytochem. 1996 Dec;44(12):1363-72
references.pmid: 8985128
references.citation: Theriogenology. 2004 Jun;61(9):1675-89
references.pmid: 15019463