title: Assisted reproduction with in-vitro-cultured testicular spermatozoa in cases of severe germ cell apoptosis: a pilot study.
abstract: BACKGROUND
  Apoptosis-related cell damage is known to compromise success rates of assisted reproduction with ejaculated spermatozoa. This study was undertaken to determine whether the frequency of apoptosis-related cell damage and reproductive performance of testicular spermatozoa from men with non-obstructive azoospermia can be improved by in-vitro culture.
  
  
  METHODS
  Testicular tissue samples were cultured for 2 days in the presence of 50 IU/l FSH and 1 micromol/l testosterone. The frequency of spermatozoa showing DNA strand breakage and plasma membrane phosphatidylserine externalization was compared in before-culture and after-culture samples. The after-culture samples were used in assisted reproduction attempts.
  
  
  RESULTS
  In a group of 11 azoospermic patients with at least two previous intracytoplasmic sperm injection (ICSI) failures, the incidence of DNA strand breakage was high in living testicular spermatozoa from before-culture samples, but significantly lower in after-culture samples (96 versus 30%, P < 0.001). The same applied to the incidence of phosphatidylserine externalization in the motile sperm subpopulation from the before-culture and after-culture samples (83 versus 6%, P < 0.001). Seven ongoing clinical pregnancies (six with fresh embryos and one with cryopreserved embryos) were established.
  
  
  CONCLUSIONS
  Severe testicular sperm apoptosis may become a new indication for testicular tissue in-vitro culture before ICSI.
authors.affiliation: Laboratoire d'Eylau, Paris, France. rtesarik@hotmail.com
authors.forename: J
authors.initials: J
authors.lastname: Tesarik
authors.forename: E
authors.initials: E
authors.lastname: Greco
authors.forename: C
authors.initials: C
authors.lastname: Mendoza
mesh_terms: D000328:Adult; D017209:Apoptosis; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D046508:Culture Techniques; D004249:DNA Damage; D004624:Embryo Transfer; D005260:Female; D006801:Humans; D008297:Male; D009845:Oligospermia / Q000628:therapy*; D010718:Phosphatidylserines / Q000032:analysis / Q000378:metabolism; D010865:Pilot Projects; D011247:Pregnancy; D011256:Pregnancy Outcome; D020554:Sperm Injections, Intracytoplasmic; D013081:Sperm Motility; D013094:Spermatozoa / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D013737:Testis / Q000473:pathology*; D020858:Tissue and Organ Harvesting
chemical_list: D010718:Phosphatidylserines