title: Long-term cultures of testicular biopsies from boys with cryptorchidism: effect of FSH and LH on the number of germ cells.
abstract: BACKGROUND
  A long-term culture system of testicular biopsies from boys with undescended testes was established to evaluate the effect of gonadotrophins on germ cell survival and growth.
  
  
  METHODS
  Biopsies from 25 boys with cryptorchidism, aged 1.1-9.9 years, were obtained at the time of surgery. Each biopsy was subdivided into fragments and cultured for 1 or 3 weeks in a testis culture medium without gonadotrophins or supplemented with FSH (50 IU/l) or LH (5 IU/l), or a combination of FSH (50 IU/l) plus LH (5 IU/l). The survival of the germ cells was evaluated by calculating the ratio of spermatogonia and gonocytes per cross-sectioned testicular tubule, the S/T ratio.
  
  
  RESULTS
  All cultured fragments maintained their overall morphology. A significantly reduced S/T ratio was observed in the fragments cultured in the presence of LH (P = 0.006), independently of the culture period. The mean tubular diameter in fragments cultured for 1 week with hormones was significantly larger than that of the controls (P < 0.0002). All cultured fragments had a larger tubular diameter than the uncultured fragments (P = 0.0002).
  
  
  CONCLUSIONS
  This culture system supports survival of spermatogonia for at least 3 weeks. LH either alone or in combination with FSH results in a significantly reduced S/T ratio.
authors.affiliation: Laboratory of Reproductive Biology, section 5712, The National University Hospital, Blegdamsvej 9, DK-2100, Copenhagen, Denmark. hplarsen@e-box.dk
authors.forename: Hans-Peter Ejler
authors.initials: HP
authors.lastname: Larsen
authors.forename: Jørgen
authors.initials: J
authors.lastname: Thorup
authors.forename: Lene Theil
authors.initials: LT
authors.lastname: Skovgaard
authors.forename: Dina
authors.initials: D
authors.lastname: Cortes
authors.forename: Anne Grete
authors.initials: AG
authors.lastname: Byskov
mesh_terms: D001706:Biopsy; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003456:Cryptorchidism / Q000473:pathology*; D003470:Culture Media / Q000494:pharmacology; D003584:Cytological Techniques; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006801:Humans; D007223:Infant; D007986:Luteinizing Hormone / Q000494:pharmacology*; D008297:Male; D012671:Seminiferous Tubules / Q000473:pathology; D013076:Sperm Count; D013094:Spermatozoa / Q000187:drug effects* / Q000473:pathology* / Q000502:physiology; D013737:Testis / Q000473:pathology*; D013997:Time Factors
chemical_list: D003470:Culture Media; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone