Epididymal spermatozoa: recovery and subsequent improvements of mouse epididymal spermatozoa via the SpermPrep filtration method.
This study was designed to determine the effects of Sephadex filtration (SpermPrep 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 degrees C for 15 min. The specimen was then split into 2 aliquots, with Alquot 1 as the control and Aliquot 2 used for filtration. SpermPrep I column was employed according 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 filtered fractions were evaluated and incubated at 37 degrees C and assessed for percentage and grade of motility (0-4) every 30 min 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 SpermPrep method improved the percentage and grade of motility (p < 0.05) but not the percentage normal morphology of the spermatozoa. Also the SpermPrep I enabled the recovery of 45% (8.3 x 10(6) spermatozoa) of the total spermatozoa processed in the control aliquot (18.4 x 10(6) spermatozoa) which is consistent with previous observations. Most importantly, filtered spermatozoa incubated for 3 hr showed greater percentage and grade of motility than the control spermatozoa (63% and 1.66 vs. 39% and 0.82, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
Zavos
P M
PM
Department of Animal Science, University of Kentucky, Lexington 40523, USA.
Correa
J R
JR
Zarmakoupis
P N
PN
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D003911:Dextrans; D004822:Epididymis / Q000166:cytology; D005374:Filtration / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D013076:Sperm Count / Q000379:methods; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology*
D003911:Dextrans; C025614:sephadex