Preparation of human frozen-thawed seminal specimens using the SpermPrep filtration method: improvements over the conventional swim-up method.
OBJECTIVE
To examine the qualitative and quantitative characteristics of frozen-thawed spermatozoa recovered through the SpermPrep (ZBL, Inc., Lexington, KY) filtration method or the swim-up technique for the purpose of intrauterine insemination (IUI) or other techniques for assisted reproduction.


METHODS
Thirty pairs of frozen specimens purchased from three commercial semen suppliers were used in this study. Each pair consisted of two aliquots from the same semen specimen.


METHODS
Spermatozoa recovered via the SpermPrep filtration and swim-up processes were evaluated for sperm numbers recovered, sperm motility, grade of sperm motility, percentage of morphologically normal spermatozoa, the response of spermatozoa to a hypo-osmotic environment (hypo-osmotic swelling test), and the amount of debris present.


RESULTS
Application of the SpermPrep filtration method resulted in recovery of significantly greater numbers of spermatozoa (P less than 0.01) than were recovered with the swim-up method (31.1 +/- 3.2 x 10(6) versus 10.2 +/- 1.8 x 10(6) spermatozoa, respectively). This represents a mean recovery of approximately one half (49%) of all spermatozoa applied to the filter, whereas for the swim-up method, it was only 15%. The overall quality of recovered spermatozoa was virtually identical between the two methods (P greater than 0.05). The percent motile sperm improved by a mean of 18% to 20%, the grade of motility improved by a mean of 0.4 points (scale 0 to 4), the percent of morphologically normal spermatozoa increased by a mean of approximately 10%, the percent of spermatozoa reactive to a hypo-osmotic medium test increased by a mean of approximately 9%, and the debris score decreased by a mean of 0.2 to 0.3 points (scale 0 to 4). Most importantly, the mean total number of motile, morphologically normal spermatozoa after filtration through the SpermPrep column was 20.2 +/- 1.1 x 10(6), representing a mean recovery of 73% of the normal spermatozoa originally applied to the column. This was 316% greater than the yield obtained with the swim-up method (6.4 +/- 0.8 x 10(6)), which was significantly greater (P less than 0.01) than that recovered via the swim-up method. Also, the time required to harvest sperm through SpermPrep filtration was 20 to 25 minutes versus 80 minutes required for the swim-up method (P less than 0.05).


CONCLUSIONS
Considering that the effectiveness of frozen-thawed semen is already limited when compared with fresh semen, SpermPrep filtration is the method of choice over the swim-up technique of sperm selection because the former provides significantly greater numbers of high quality sperm. It should be considered as an adjunct in semen preparation for IUI or other forms of assisted reproduction.
Zavos
P M
PM
Andrology Institute of Lexington, Kentucky.
D005069:Evaluation Studies as Topic; D005374:Filtration; D005615:Freezing; D006801:Humans; D008297:Male; D012661:Semen; D013048:Specimen Handling / Q000379:methods*; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology