title: Optimal use of fresh and frozen-thawed testicular sperm for intracytoplasmic sperm injection in azoospermic patients.
abstract: OBJECTIVE
  To optimize the use of fresh and frozen-thawed testicular biopsy specimens from patients with azoospermia.
  
  
  METHODS
  Fifty-one patients suffering from obstructive and non-obstructive azoospermia underwent testicular sperm extraction (TESE). The specimens were divided and used for either in vitro maturation or freezing for a future intracytoplasmic sperm injection (ICSI) cycle.
  
  
  RESULTS
  At initial testicular sperm extraction, very few motile spermatozoa were seen. After 24 h of in vitro maturation, sperm motility increased remarkably, with a maximum motility rate seen between 48 and 72 h of culture. Motile spermatozoa were observed up to 120 h in culture. In the 22 fresh ICSI cycles, a total of 294 oocytes were injected using motile sperm and 212 oocytes demonstrated normal 2PN formation (fertilization rate, 72.1%). In 36 frozen-thawed ICSI cycles, a total of 454 oocytes were injected and 302 oocytes became 2PN (66.5%). On day 3, high quality embryos were observed in 54.2% of fresh cycles and 54.1% of frozen cycles (P > 0.05). The clinical pregnancy rate did not show a significant difference between using fresh (59%) and frozen (55.5%) testicular biopsy sperm for ICSI (P > 0.05), but the embryo implantation rates did differ significantly between fresh (29.5%) and frozen-thawed (22.2%) cycles (P < 0.05). A total of 33 healthy babies have been born from 22 women, giving birth after 58 embryo transfer attempts (38%).
  
  
  CONCLUSIONS
  The freezing and in vitro culturing of testicular biopsy tissue is a very reliable approach for the management of testicular biopsy specimens from azoospermic patients, and offers the possibility of several treatments of IVF/ICSI from a single sample.
authors.affiliation: Arizona Center for Reproductive Endocrinology and Infertility, 5190 E Farness Drive, #114, Tucson, AZ, 85712, USA. bwu13@yahoo.com
authors.forename: Bin
authors.initials: B
authors.lastname: Wu
authors.forename: Diana
authors.initials: D
authors.lastname: Wong
authors.forename: Suzhen
authors.initials: S
authors.lastname: Lu
authors.forename: Stephen
authors.initials: S
authors.lastname: Dickstein
authors.forename: Marta
authors.initials: M
authors.lastname: Silva
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Gelety
mesh_terms: D000328:Adult; D001706:Biopsy; D015925:Cryopreservation; D046508:Culture Techniques; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009845:Oligospermia; D011247:Pregnancy; D018873:Pregnancy Rate; D020554:Sperm Injections, Intracytoplasmic; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology