IMPROVEMENTS AND SHORT-TERM VIABILITY OF MOUSE EPIDIDYMAL SPERMATOZOARECOVERED THROUGH THE SPERMPREP(TM) FILTRATION METHOD

This study was designed to determine the effects of Sephadex filtratio n (Spermprep(TM)I method) on the separation of motile, morphologically normal, mouse epididymal spermatozoa and to study the viability of th e recovered spermatozoa over a 3-h incubation period. Spermatozoa were harvested from the caudae epididymie (5 animals per run or replicatio n; n=10) following bilateral testicular excision, after which they wer e incubated in 2-ml of Test-Yolk buffer (TYB) at 37 degrees C for 15-m in. The specimens were then split into 2 1-ml aliquots, with Aliquot 1 as the control and Aliquot 2 as the filtered sample. The Spermprep(TM )I column was employed according to the manufacturer's specifications using TYB. During filtration (10-min), 2 different fractions were obta ined: first 5-min (Sample 1) and second 5-min (Sample 2). The 2 fracti ons were evaluated and incubated at 37 degrees C and assessed for perc entage of motility and grade of motility (0 to 4) every 30-min for 3-h . Filtration resulted in a significant improvement in the percentage a nd grade of motility (91.5% and 3.0 vs 76.5% and 2.5, respectively). T he results indicate that filtration with the Spermprep(TM)I method imp roved the percentage and grade of motility (P<0.05) but not the percen tage of normal morphology of the spermatozoa. In addition, the Spermpr ep(TM)I method enabled the recovery of 45% (8.3 x 10(6) spermatozoa re covered) of the total number of spermatozoa processed in the control a liquot (18.4 x 10(6) spermatozoa), which is consistent with previous o bservations. Most importantly, filtered spermatozoa incubated for 3-h showed a greater percentage and grade of motility than the control spe rmatozoa (63% and 1.66 vs 39% and 0.82, respectively. The Spermprep(TM )I filtration method selected a higher proportion of quality spermatoz oa, which also displayed significant long-term motility (longevity) du ring in vitro incubation.