Background: To compare four different spermatozoa preparation methods in semen samples with respect to recovery rate, percent motility, path and progressive velocity, and nitric oxide (NO) production before and after centrifugation.
Methods: Each of 36 semen specimens was diluted to 1 x 10(6)/ml and divided into four 1-ml aliquots for sperm processing using four methods: swim-up, two (Percoll II) and three (Percoll III) layer Percoll (Pharmacia Biotech AB, Uppsala, Sweden) gradients and albumin columns. The motile sperm recovery rate, percent motility and motion parameters were measured for each semen specimen (n = 36) before and after processing using the four methods. NO was measured with the use of a chemiluminescence method before and after centrifugation (n = 17).
Results: The sperm recovery rate was higher using both Percoll gradients than with the other two methods (Percoll II, 68 +/- 20.5%, Percoll III, 75.3 +/- 22.2% vs swim-up, 25.8 +/- 9.9% and albumin, 33.1 +/- 20.7%). The results were similar for total motile cells (Percoll II, 3.55 +/- 1.72 x 10(6)/ml, Percoll III, 4.25 +/- 1.29 x 10(6)/ml vs swim-up, 1.19 +/- 0.71 x 10(6)/ml and albumin, 1.89 +/- 1.36 x 10(6)/ml). Both Percoll methods and the albumin method produced a statistically significant improvement over the swim-up method in motility. The albumin column method resulted in the least path and progressive velocities (51.3 +/- 15.4 microns/s and 46.2 +/- 16.5 microns/s, respectively). NO produced during sperm processing did not differ significantly among the four processing methods (swim-up, 4,531 +/- 1,626 nM, Percoll II, 5,119 +/- 3,969 nM, Percoll III, 6,060 +/- 5,512 nM, albumin 4,838 +/- 2,462 nM).
Conclusions: Our data suggest that Percoll methods are superior to swim-up and albumin columns for sperm preparation, yielding good sperm recovery, motility and motion characteristics. The trend toward lower NO levels among samples prepared using albumin columns did not reach statistical significance.