Treatment of freeze-thawed testicular sperms with Human Follicular Fluid (HFF) improves their motility and is useful to select viable sperms for ICSI procedure

Abstract

Objective: The motiltiy of sperms obtained from testicular biopsy is generally very low.Since in some cases, it is nessesary to freeze the biopsied tissue in order to prevent of repeated testis biopsy, and for future use of testicular sperms in ICSI,on the other hand, many of these cells may lose their viability during cryopreservation, it is reasonable to find a way which may help us to select viable sperms after thawing, for injecting them into oocytes. According to previous studies on the positive effects of hFF on sperm motility and its capacitation,we tried to take advantage of this property and to evaluate the motility of freeze-thawed testicular sperms, following culture in different combinations of hFF and Ham’s F-10. Design: Comparison of testicular sperm motility immediately after thawing,and following culture in different concentrations of hFF. Materials and Methods: The biopsied testicular tissue (n=8) was subjected to mechanical mincing by two glass slides and recovered testicular sperms were freezed with conventional sperm freeze solution (glycerol 15%). After thawing, samples were allocated to four groups as follow: Group 1(Control);Ham’s F-10/without culture,Group 2; Ham’s F-10+0% hFF/with 24 hrs culture, Group 3; Ham’s F-10 +25% hFF/with 24hrs culture, Group 3; Ham’s F-10 +50% hFF/with 24hrs culture.Culture was done under 5% CO2 and 37° C .In group 1 (Control), motility was evaluated immediately after thawing .In Group 2,evaluation was done to determine if culture alone has a positive effect on motility and in remainig groups(3 and 4), the effect of different concentrations of hFF on testicular sperm motility was determined. At least 100 sperms were counted for motiltiy scoring in each group. Statistical analysis was performed by paired t-test between control and experimental groups,and a comparison of Mean+SD for motility between groups 2,3 and 4 was done to define the best concentration of hFF for testicular sperm culture after thawing. P<0.05 considered significant. Results: Sperm motility immediately after thawing was 3.00+ 4.40%(Group 1).After 24 hrs of culture, motility was 21.25+12.61% (Ggroup2),30.75+16.23% (Group 3) and 43.0+17.00% (Group 4) respectively.The differences between control and other groups(cultured in FF) was statistically significant. Also the motility was significantly higher in group 4 compared with other groups(2,3). Conclusion: These results suggest that 24 hrs culture of freeze-thawed testicular sperms in media containing 50% hFF may improve its motility,and, in addition of helping us to select viable sperms for ICSI, may be beneficial for these cells in aspect of optimizing their quality and ability to fertilize oocytes.