title: In vitro culture facilitates the selection of healthy spermatids for assisted reproduction.
abstract: OBJECTIVE
  To evaluate the potential usefulness of in vitro culture of germ cells for distinguishing between healthy and apoptotic spermatids.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  Private assisted reproduction laboratories and a university department.
  
  
  METHODS
  Men with secretory azoospermia who were candidates for assisted reproductive treatment.
  
  
  METHODS
  Testicular biopsy samples were cultured in the presence of FSH and testosterone for 48 hours. Germ cell apoptosis before and after culture was evaluated by terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling.
  
  
  METHODS
  The percentage of germ cells at different stages of spermatogenesis that showed apoptosis-related DNA damage.
  
  
  RESULTS
  In fresh samples, high levels of apoptosis were detected at those stages of spermatogenesis at which major developmental blocks occurred, with a maximum at the most advanced stage detected. In contrast, apoptotic cells were considerably less well represented at the most advanced stage after culture.
  
  
  CONCLUSIONS
  In addition to the previously described facilitation of spermatid recognition and the progression of cytoplasmic maturation, in vitro culture of germ cells is useful to overcome the danger of inadvertent use of apoptotic spermatids for assisted reproduction.
authors.affiliation: Laboratoire d'Eylau, Paris, France. cmendoza@goliat.ugr.es
authors.forename: J
authors.initials: J
authors.lastname: Tesarik
authors.forename: C
authors.initials: C
authors.lastname: Mendoza
authors.forename: E
authors.initials: E
authors.lastname: Greco
mesh_terms: D017209:Apoptosis / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology; D006801:Humans; D008297:Male; D008540:Meiosis / Q000502:physiology; D009845:Oligospermia / Q000473:pathology*; D011446:Prospective Studies; D012099:Reproductive Techniques; D013087:Spermatids / Q000166:cytology*; D013094:Spermatozoa / Q000166:cytology*