Increased sperm motility after in-vitro culture of testicular biopsies from obstructive azoospermic patients results in better post-thaw recovery rate

Abstract

The objective of this study was to optimize the use of testicular biopsies in 14 patients with obstructive azoospermia. Testicular specimens were retrieved from six patients (group I) and cultured at 32 and 37 degrees C for up to 20 days; changes in percentage motile spermatozoa were compared. In four men of group I, one portion of the specimen was frozen at retrieval, and changes in post-thaw motility after 24 h of culture at 37 degrees C were recorded. In the other eight patients (group II), testicular specimens were frozen at retrieval and after 72 h culture at 37 degrees C. Pre and post-freezing motility and post-thaw recovery rate were compared. No significant differences were observed until day 8 in the improvement of motility between 32 and 37 degrees C in-vitro culture. Maximum motility was reached, under both conditions, between 48 h and 72 h. Post-thaw 24 h culture at 37 degrees C of specimens frozen at retrieval did not improve motility; however, 72 h pre-freezing culture significantly improved initial motility (P: < 0.01), post-thaw motility (P: < 0.01) and post-thaw recovery rate (P: < 0. 001). The higher recovery rate of samples frozen 3 days after retrieval allows more economical use of the tissue that is available.

Clinical freeze preservation of whole human testicular tissue: the practicality of pre-freeze in vitro culture (IVC) to optimize post-thaw sperm motility

“…We have observed the latter phenomenon in this and prior studies [5], observing a downward shift in the initial % post-thaw motility in each movement category. The IVC outcomes of this study are consistent with prior observations [5,8,9]. The more overt the movement of sperm, the easier the task at hand is for the Embryologist diligently assessing the cellular milleu to isolate viable sperm for injection.…”

“…Initially, 5-10% twitching may be observed in a fresh specimen (0-3 hours), with the infrequent occurrence of more active sperm. It has been shown that in vitro culture (IVC) of testicular tissue does promote motility onset and progression [6,7], with an intermediate incubation temperature of 30-32ºC optimizing motility formation and longevity [5,8,9]. Although testicular sperm motility is not required to achieve fertilization by fresh sperm post-ICSI [10], it is generally considered preferential for increasing pregnancy potential [11].…”

Optimal use of fresh and frozen-thawed testicular sperm for intracytoplasmic sperm injection in azoospermic patients

“…China. 4 To whom correspondence should be addressed; e-mail: bwu13@ yahoo.com. sperm for ICSI is now a widely practiced technique in the treatment of men with diagnosed both obstructive and non-obstructive azoospermia.…”

“…The optimal use of testicular biopsies for these treatments would therefore reduce surgical extractions to retrieval of a single sample. In the past few years, many groups have suggested methods to optimize the use of testicular biopsy specimens (2)(3)(4)(5)(6). Several strategies have been developed to optimally manage a testicular biopsy, including improvement of sperm motility and increasing the motile sperm recovery rate after freezing.…”

Viable pregnancies can occur after 24-hour incubation of ejaculated sperm before intracytoplasmic sperm injection

“…Longer incubations are often necessary with spermatozoa retrieved from testicular biopsy or epididymal aspiration, presumably to stimulate in vitro maturation of the immature testicular sperm (1)(2)(3). Incubation of testicular sperm showed improved sperm motility, which peaked at 48 hours of in vitro culture (4,5). Pregnancies using testicular sperm incubated for >48 hours have been reported (6,7).…”