Sperm Separation Technique Using Iodixanol Compare to Percoll Gradient

Abstract

Objective: To evaluate the use of iodixanol for the separation of sperm motile fraction and the effect

on sperm morphology comparing to Percoll gradient.           

Methods: Thirty semen samples from infertile patients were studied. The initial evaluation of sperm
count and morphology was performed. The samples were then equally aliquoted into 2 parts and were
separately prepared by discontinuous gradient using Percoll and iodixanol. The yielded sperm from the
two preparations were compared under the following parameters: motile and non-motile sperm count,
progressive motion and percentage of normal morphology.
Results: Concentrations of motile sperm after sperm preparation by both Percoll and iodixanol were
comparable (17.4+11.7 VS 16.3+14.0´ 106/ml). Non-motile sperm count from Percoll and iodixanol
were lower than the initial sperm (6.2+9.9, 18.1+19.1, and 24.2+15.9 ´ 106/ml, respectively). Iodixanol
produced higher non-motile sperm concentration compared to Percoll. Both gradient materials gave
comparable improvement of progressive movement from 2.2+0.3 in initial sperm to 2.9+0.2 by Percoll
and 2.8+0.2 by iodixanol. Percoll gradient gave a significant improvement of the percentage of normal
sperm morphology from 14.1+3.3 % in initial sperm to 16.8+5.3 %, while there was no change after
iodixanol gradient. Percentages of head and tail defects were not changed after preparation by either
gradient but midpiece defect was significantly decreased from 11.1+3.6 % to 7.8+4.3 % after Percoll
process.
Conclusion: Although motile sperm yielded by Percoll and iodixanol were comparable, Percoll
gradient gave a better percent motility because it could separate more non-motile sperm. Percoll
gradient also gave a better percentage of morphologically normal spermatozoa compared to iodixanol
gradient. This new medium could replace Percoll if some preparation procedures are adjusted.