抄録
ZAVOS, P.M, CORREA, J.R. and ZARMAKOUPIS, P.N. Epididymal Spermatozoa: Recovery and Subsequent Improvements of Mouse Epididymal Spermatozoa via the SpermPrepTM Filtration Method. Tohoku J. Exp. Med., 1995, 175 (2), 101-109 -This study was designed to determine the effects of Sephadex filtration (SpermPrepTM method) on the separation of viable motile, morphologically normal mouse epididymal spermatozoa and to study the viability of the recovered spermatozoa over a 3 hr incubation period. Spermatozoa were harvested from the caudae epididymides (5 animals per run or replication; n=10) following bilateral testicular excision and incubated in 2-ml of Test-Yolk buffer (TYB) at 37°C for 15 min. The specimen was then split into 2 aliquots, with Alquot 1 as the control and Aliquot 2 used for filtration. SpermPrepTMI column was employed according to the manufacturer's specifications (ZBL, Inc., Lexington, KY, USA) using TYB. During filtration (10min), different fractions were obtained: first 5min (Sample 1) and second 5min (Sample 2). The filtered fractions were evaluated and incubated at 37°C and assessed for percentage and grade of motility (0-4) every 30min for 3 hr. Filtration resulted in a significant improvement in percentage and grade of motility (91.5% and 3.0 vs. 76.5% and 2.5, respectively). The results point out very clearly that the filtration via the SpermPrepTM method improved the percentage and grade of motility (p<0.05) but not the percentage normal morphology of the spermatozoa. Also the SpermPrepTMI enabled the recovery of 45% (8.3×106 spermatozoa) of the total spermatozoa processed in the control aliquot (18.4×106 spermatozoa) which is consistent with previous observations. Most importantly, filtered spermatozoa incubated for 3hr showed greater percentage and grade of motility than the control spermatozoa (63% and 1.66 vs. 39% and 0.82, respectively). This is of significant importance since the filtration via the SpermPrepTMI has selected not only greater quality spermatozoa but also spermatozoa with significantly long-term motility (longevity) during in vitro incubation.
