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 (percentage motile and progressively motile) of freshly collected testicular and epididymal spermatozoa and following freeze/thaw of the same spermatoz oa from a man with obstructive azoospermia. Washed spermatozoa were cu ltured in micro droplets under paraffin oil or in test tubes using HEP ES-buffered or bicarbonate-buffered medium containing 10% human serum. In fresh testicular sperm cultures 60-65% of the sperm cells became m otile within 2 days of culture; the motility was maintained for a furt her 4-5 days before a decline was observed. The progressive motility i mproved 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 maintaine d for only 2-3 days before it declined. Furthermore, only 10-12% of th e 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 im motile in both freshly collected and frozen/thawed spermatozoa. All cu lture systems supported sperm motility. It is clear that testicular sp ermatozoa, particularly from men with obstructive azoospermia, can be collected and maintained in vitro for up to 1 week before the oocyte r etrieval but when frozen testicular or epididymal spermatozoa are used it is more reliable to thaw these spermatozoa on the day of intracyto plasmic sperm injection.