EPIDIDYMAL SPERMATOZOA - RECOVERY AND SUBSEQUENT IMPROVEMENTS OF MOUSE EPIDIDYMAL SPERMATOZOA VIA THE SPERMPREP(TM) FILTRATION METHOD

This study was designed to determine the effects of Sephadex filtratio n (SpermPrep(TM) method) on the separation of viable motile, morpholog ically normal mouse epididymal spermatozoa and to study the viability of the recovered spermatozoa over a 3 hr incubation period. Spermatozo a were harvested from the caudae epididymides (5 animals per run or re plication; n=10) following bilateral testicular excision and incubated in 2-ml of Test-Yolk buffer (TYB) at 37 degrees C for 15 min. The spe cimen was then split into 2 aliquots, with Alquot 1 as the control and Aliquot 2 used for filtration. SpermPrep(TM)I column was employed acc ording to the manufacturer's specifications (ZBL, Inc., Lexington, KY, USA) using TYB. During filtration (10 min), different fractions were obtained: first 5 min (Sample 1) and second 5 min (Sample 2). The filt ered fractions were evaluated and incubated at 37 degrees C and assess ed for percentage and grade of motility (0-4) every 30 min for 3 hr. F iltration resulted in a significant improvement in percentage and grad e of motility (91.5% and 3.0 vs. 76.5% and 2.5, respectively). The res ults point out very clearly that the filtration via the SpermPrep(TM) method improved the percentage and grade of motility (p<0.05) but not the percentage normal morphology of the spermatozoa. Also the SpermPre p(TM)I enabled the recovery of 45% (8.3 x 10(6) spermatozoa) of the to tal spermatozoa processed in the control aliquot (18.4 x 10(6) spermat ozoa) which is consistent with previous observations. Most importantly , filtered spermatozoa incubated for 3 hr showed greater percentage an d 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 SpermPrep(TM)I has selected not only greater qualit y spermatozoa but also spermatozoa with significantly long-term motili ty (longevity) during in vitro incubation.