title: Effect of antifreeze protein supplementation in vitrification medium on mouse oocyte developmental competence.
abstract: OBJECTIVE
  To investigate the effect of antifreeze protein (AFP) supplementation during mouse oocyte vitrification on the survival, fertilization and embryonic development.
  
  
  METHODS
  Animal study.
  
  
  METHODS
  University laboratory.
  
  
  METHODS
  BDF-1 mice.
  
  
  METHODS
  In vivo-matured metaphase II oocytes were vitrified with the use of CryoTop by two-step exposure to equilibrium and vitrification solution supplemented or not with 500 ng/mL AFP III.
  
  
  METHODS
  Postwarming survival, fertilization, embryonic development up to blastocyst in vitro, morphology of spindle and chromosome, membrane integrity, adenosine triphosphate (ATP) contents, and several gene expressions.
  
  
  RESULTS
  In the AFP-treated group, blastocyst formation rate was significantly higher and blastomere count with positive caspase was significantly lower compared with the nontreated group. Rate of intact spindle/chromosome, stable membrane, and ATP contents were significantly higher in AFP group. AFP group showed higher Mad2 and lower Eg5 gene expression. Both vitrification groups showed increased Hsf1, Zar1, and Zp1/Zp2 expression and decreased Hook1 and Zp3 expression compared with fresh control samples.
  
  
  CONCLUSIONS
  Supplementation of AFP in vitrification medium has a protective effect on mouse oocytes for chilling injury; it can preserve spindle/membrane integrity and intracellular ATP contents. More stable spindle integrity in the AFP group may be associated with higher Mad2 and lower Eg5 gene expression.
authors.affiliation: Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Gyeonggi, South Korea.
authors.forename: Jun Woo
authors.initials: JW
authors.lastname: Jo
authors.forename: Byung Chul
authors.initials: BC
authors.lastname: Jee
authors.forename: Jung Ryeol
authors.initials: JR
authors.lastname: Lee
authors.forename: Chang Suk
authors.initials: CS
authors.lastname: Suh
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D021301:Antifreeze Proteins / Q000494:pharmacology*; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051379:Mice; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D058989:Vitrification
chemical_list: D021301:Antifreeze Proteins; D003451:Cryoprotective Agents; D000255:Adenosine Triphosphate