title: Vitrification of mouse embryos with super-cooled air.
abstract: OBJECTIVE
  To develop a closed vitrification device (i.e., one that requires no direct contact with liquid nitrogen) for successful cryostorage of embryos.
  
  
  METHODS
  Prospective laboratory research study.
  
  
  METHODS
  University-based research laboratory.
  
  
  METHODS
  F1 mice and mouse embryos.
  
  
  METHODS
  Mouse embryos were vitrified using two methods and compared with nonvitrified controls. Embryos were vitrified on a device by either [1] presealing it within a straw before plunging into liquid nitrogen or [2] placing the straw into liquid nitrogen so that the air inside the straw is super-cooled before inserting the device holding the embryos.
  
  
  METHODS
  Survival, subsequent embryo development, and cell number were determined. Embryos were also cryopreserved for 12 months to assess long-term storage. Synchronized ETs were performed to compare viability with nonvitrified control embryos.
  
  
  RESULTS
  All embryos survived with both techniques. Day-4 and -5 embryo development was comparable between the two vitrification methods. Use of the presealing method resulted in a significantly lower mean cell number than the postsealing method and control. Long-term storage did not affect subsequent embryo development or cell number. The implantation and fetal development rates of embryos vitrified with super-cooled air were comparable to those for nonvitrified control embryos.
  
  
  CONCLUSIONS
  These data demonstrate that a closed vitrification device (Rapid-i), which does not require direct liquid nitrogen contact for vitrification, is appropriate for vitrification and long-term storage of mouse embryos.
authors.affiliation: Department of Zoology, University of Melbourne, Melbourne, Victoria, Australia. mlarman@unimelb.edu.au
authors.forename: Mark G
authors.initials: MG
authors.lastname: Larman
authors.forename: David K
authors.initials: DK
authors.lastname: Gardner
mesh_terms: D000388:Air; D000818:Animals; D003080:Cold Temperature; D004624:Embryo Transfer / Q000379:methods; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice / Q000196:embryology*; D011247:Pregnancy; D011446:Prospective Studies; D058989:Vitrification