Case report: 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

The present report describes the motility changes in vitro (percentagemotile and progressively motile) of freshly collected testicularand epididymal spermatozoa and following freeze/thaw of thesame spermatozoa from a man with obstructive azoospermia. Washedspermatozoa were cultured in micro droplets under paraffin oilor in test tubes using HEPES-buffered or bicarbonate-bufferedmedium containing 10% human serum. In fresh testicular spermcultures 60–65% of the sperm cells became motile within2 days of culture; the motility was maintained for a further4–5 days before a decline was observed. The progressivemotility unproved markedly on the third day of culture and itpeaked around day 5. Only a small number of frozen/ thawed testicularspermatozoa became motile during in-vitro culture (15–20%)and the motility was maintained for only 2–3 days beforeit declined. Furthermore, only 10–12% of the spermatozoashowed progressive motility. Spermatozoa recovered from micro-epididymalsperm aspiration (MESA) showed a gradual decrease in progressivemotility and in 5 days all sperm cells were found to be immotilein both freshly collected and frozen/thawed spermatozoa. Allculture systems supported sperm motility. It is clear that testicularspermatozoa, particularly from men with obstructive azoospermia,can be collected and maintained in vitro for up to 1 week beforethe oocyte retrieval but when frozen testicular or epididymalspermatozoa are used it is more reliable to thaw these spermatozoaon the day of intracytoplasmic sperm injection.