title: Can prepubertal human testicular tissue be cryopreserved by vitrification?
abstract: OBJECTIVE
  To assess vitrification of prepubertal human testicular tissue in vitro.
  
  
  METHODS
  Case report.
  
  
  METHODS
  Academic research unit.
  
  
  METHODS
  Two patients (6 and 12 years of age) who were to start gonadotoxic treatment for chronic granulomatous disease and acute lymphoblastic leukemia.
  
  
  METHODS
  Long-term (10-day) organotypic culture performed immediately after vitrification and warming. Fresh tissue and tissue cryopreserved by slow-freezing were used as control samples.
  
  
  METHODS
  Spermatogonial cell survival (MAGE-A4) and proliferation (Ki67) were evaluated by immunohistochemistry (IHC) and tubular integrity by light microscopy.
  
  
  RESULTS
  Qualitative analysis revealed that histologic characteristics of spermatogonia and Sertoli cells were preserved, as were cell-cell cohesion and cell adhesion to the basement membrane, in vitrified tissue as well as in frozen and fresh control samples. Survival of spermatogonia and their ability to proliferate as evidenced by IHC was also confirmed in cultured fresh, slow-frozen, and vitrified tissue.
  
  
  CONCLUSIONS
  Vitrification, having the advantage of being a faster and more convenient method, shows promise as an alternative strategy to slow-freezing in the emerging field of immature testicular tissue cryopreservation.
authors.affiliation: Department of Gynecology, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Mara
authors.initials: M
authors.lastname: Curaba
authors.forename: Jonathan
authors.initials: J
authors.lastname: Poels
authors.forename: Anne
authors.initials: A
authors.lastname: van Langendonckt
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
authors.forename: Christine
authors.initials: C
authors.lastname: Wyns
mesh_terms: D002478:Cells, Cultured; D002648:Child; D015925:Cryopreservation / Q000379:methods*; D005240:Feasibility Studies; D006105:Granulomatous Disease, Chronic / Q000628:therapy; D006801:Humans; D007248:Infertility, Male / Q000517:prevention & control; D008297:Male; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000628:therapy; D011627:Puberty / Q000502:physiology; D055101:Semen Analysis; D013737:Testis; D058989:Vitrification