title: Cryopreservation of prepubertal mouse testicular tissue by vitrification.
abstract: OBJECTIVE
  To compare cryopreservation of prepubertal testicular tissue after vitrification (V) and slow-freezing (SF).
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  Academic research unit.
  
  
  METHODS
  Six-day-old mice.
  
  
  METHODS
  After cryopreservation, viability tests (n = 10) and short-term culture (1 and 3 days) (n = 5) were performed. A comparison was made with fresh (FR) and noncultured controls (FR Ctrl).
  
  
  METHODS
  Tissue viability was assessed by lactate dehydrogenase release assay. Apoptosis (caspase-3) and proliferation (Ki67) were evaluated by immunohistochemistry, and tubular diameter, integrity, and cell density by light microscopy.
  
  
  RESULTS
  Lactate dehydrogenase release was greater after SF than V (54.6% vs. 26.7%), whereas the mean number of apoptotic cells/tubule was higher after V than SF (2.13 vs. 0.07). On day 1, a decrease in cell density was noted in both cryopreserved groups, but this difference was not subsequently observed. On day 3, an increase in proliferation was seen in the SF and V groups versus FR tissue, and similar tubular diameter, integrity, and cell density were found in all cultured groups.
  
  
  CONCLUSIONS
  This study shows that both SF and V protocols preserve survival, development, and integrity of prepubertal mouse testicular tissue in short-term organotypic culture. Additional investigation should now be conducted to assess tissue functionality.
authors.affiliation: Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Mara
authors.initials: M
authors.lastname: Curaba
authors.forename: Magali
authors.initials: M
authors.lastname: Verleysen
authors.forename: Christiani Andrade
authors.initials: CA
authors.lastname: Amorim
authors.forename: Marie-Madeleine
authors.initials: MM
authors.lastname: Dolmans
authors.forename: Anne
authors.initials: A
authors.lastname: Van Langendonckt
authors.forename: Outi
authors.initials: O
authors.lastname: Hovatta
authors.forename: Christine
authors.initials: C
authors.lastname: Wyns
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
mesh_terms: D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D009924:Organ Culture Techniques; D011446:Prospective Studies; D013737:Testis; D014022:Tissue Survival / Q000502:physiology; D058989:Vitrification
chemical_list: D007770:L-Lactate Dehydrogenase