title: Orthotopic grafting of cryopreserved prepubertal testicular tissue: in search of a simple yet effective cryopreservation protocol.
abstract: OBJECTIVE
  To investigate whether solid-surface vitrification (SSV) is an effective cryopreservation strategy regarding the integrity and function of prepubertal mouse testicular tissue.
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  Academic research unit.
  
  
  METHODS
  Mice.
  
  
  METHODS
  Testicular tissue from 5- to 10-day-old GFP(+) mice was cryopreserved with the use of a conventional uncontrolled slow freezing (USF) technique and SSV before intratesticular grafting in busulfan-treated GFP(-) mice.
  
  
  METHODS
  Ultrastructural cryoinjury to spermatogonial stem cells (SSCs) and somatic cells was assessed by electron microscopy. Tubular structure was evaluated by histology, and graft survival and spermatogenic recovery by immunohistochemistry.
  
  
  RESULTS
  The tubular morphology and the proportion of ultrastructural cryodamage were similar between vitrified and slow-frozen testicular fragments. Allografting of tissue after both USF and SSV resulted in a recovery of spermatogenesis similar to fresh samples.
  
  
  CONCLUSIONS
  SSV resulted in success rates similar to USF in maintaining testicular cell ultrastructure, tubular morphology, and tissue function. These data provide further evidence that vitrification, being an inexpensive and simple technique, can be considered as an alternative for cryopreservation of prepubertal testicular tissue.
authors.affiliation: Biology of Testis, Research Laboratory for Embryology and Genetics, Vrije Universiteit Brussel, Brussels, Belgium. yoni.baert@vub.ac.be
authors.forename: Yoni
authors.initials: Y
authors.lastname: Baert
authors.forename: Ellen
authors.initials: E
authors.lastname: Goossens
authors.forename: Dorien
authors.initials: D
authors.lastname: van Saen
authors.forename: Liang
authors.initials: L
authors.lastname: Ning
authors.forename: Peter
authors.initials: P
authors.lastname: in't Veld
authors.forename: Herman
authors.initials: H
authors.lastname: Tournaye
mesh_terms: D000818:Animals; D002066:Busulfan; D015925:Cryopreservation; D004195:Disease Models, Animal; D059247:Fertility Preservation / Q000009:adverse effects / Q000379:methods*; D006085:Graft Survival; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D007150:Immunohistochemistry; D007248:Infertility, Male / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000601:surgery*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D046529:Microscopy, Electron, Transmission; D009926:Organ Preservation / Q000009:adverse effects; D020127:Recovery of Function; D046468:Sexual Development; D013091:Spermatogenesis; D013737:Testis / Q000293:injuries / Q000378:metabolism / Q000503:physiopathology / Q000637:transplantation* / Q000648:ultrastructure; D013997:Time Factors; D058989:Vitrification
chemical_list: D049452:Green Fluorescent Proteins; D002066:Busulfan