title: Changes in motility patterns during in-vitro culture of fresh and frozen/thawed testicular and epididymal spermatozoa: implications for planning treatment by intracytoplasmic sperm injection.
abstract: The present report describes the motility changes in vitro (percentage motile and progressively motile) of freshly collected testicular and epididymal spermatozoa and following freeze/thaw of the same spermatozoa from a man with obstructive azoospermia. Washed spermatozoa were cultured in micro droplets under paraffin oil or in test tubes using HEPES-buffered or bicarbonate-buffered medium containing 10% human serum. In fresh testicular sperm cultures 60-65% of the sperm cells became motile within 2 days of culture; the motility was maintained for a further 4-5 days before a decline was observed. The progressive motility improved markedly on the third day of culture and it peaked around day 5. Only a small number of frozen/thawed testicular spermatozoa became motile during in-vitro culture (15-20%) and the motility was maintained for only 2-3 days before it declined. Furthermore, only 10-12% of the spermatozoa showed progressive motility. Spermatozoa recovered from micro-epididymal sperm aspiration (MESA) showed a gradual decrease in progressive motility and in 5 days all sperm cells were found to be immotile in both freshly collected and frozen/thawed spermatozoa. All culture systems supported sperm motility. It is clear that testicular spermatozoa, particularly from men with obstructive azoospermia, can be collected and maintained in vitro for up to 1 week before the oocyte retrieval but when frozen testicular or epididymal spermatozoa are used it is more reliable to thaw these spermatozoa on the day of intracytoplasmic sperm injection.
authors.affiliation: PIVET Medical Centre, Leederville, Perth, Australia.
authors.forename: W R
authors.initials: WR
authors.lastname: Edirisinghe
authors.forename: S M
authors.initials: SM
authors.lastname: Junk
authors.forename: P L
authors.initials: PL
authors.lastname: Matson
authors.forename: J L
authors.initials: JL
authors.lastname: Yovich
mesh_terms: D000328:Adult; D002478:Cells, Cultured; D015925:Cryopreservation; D004822:Epididymis / Q000166:cytology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D006801:Humans; D008297:Male; D008845:Microinjections; D008875:Middle Aged; D013081:Sperm Motility; D013737:Testis / Q000166:cytology*; D014659:Vasectomy