Comparative study of testicular sperm maturation in vitro using ham's f10, earle's and ferticult culture media

ABSTRACT

Testicular sperm extraction [TESE] from azoospermic males followed by intracytoplasmic sperm injection [ICSI] is a recent advance in treatment of male infertility. This study describes the total number and motility changes in vitro of freshly extracted testicular sperm and compares between Ham's F-10, Earle's, and Ferticult culture media. Testicular biopsies were obtained from azoospermic patients [n=11] and processed to obtain a cell suspension that was incubated in equal portions in each culture medium supplemented with 10% human albumin for 5 days. The number of total and motile spermatozoa was evaluated on day 0 and daily afterwards for 5 days. Our results revealed that all patients were suffering from obstructive azoospermia with mean age of 39 +/- 3.36 and duration of infertility of 9.25 +/- 2.98 years. The total number of spermatozoa showed no significant changes in all days of incubation and between all used media [p>0.05]. However, the number of motile spermatozoa was significantly higher in Ferticult medium when compared with Ham's F10 and Earle's media [p <0.05]. In all used media, spermatozoa become motile within 24 hours of culture and the motility was maintained for the 4th day before a decline to the near 0 level is observed on the 5th day of incubation. Furthermore, the number of progressively motile sperm significantly peaked on day 3 of incubation in all media but the best significant yield was observed in Ferticult medium [p <0.05]. No significant correlation was found with age of patients or duration of infertility [p> 0.05]. From our study we recommend that TESE is carried out approximately 3 days before oocyte retrieval and that Ferticult medium is the medium of choice to use in in vitro cultivation for obtaining the most mature of testicular tissue sperm to be used for IVF related procedures, e.g. ICSI