Pm. Zavos et al., IMPROVEMENTS AND SHORT-TERM VIABILITY OF MOUSE EPIDIDYMAL SPERMATOZOARECOVERED THROUGH THE SPERMPREP(TM) FILTRATION METHOD, Theriogenology, 42(6), 1994, pp. 1035-1042
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.