On the use of paramagnetic beads and ferrofluids to assess and eliminate the leukocytic contribution to oxygen radical generation by human sperm suspensions.
OBJECTIVE
To develop a methodology to determine a) the leukocytic contribution to reactive oxygen species generation by human sperm suspensions and b) the therapeutic value of removing these cellular contaminants.


METHODS
Leukocytes were removed with paramagnetic beads or colloidal ferrofluids coated with anti-CD45 antibody. The sperm suspensions were monitored for oxidant generation by chemiluminescence, leukocyte contamination by immunocytochemistry, and fertilizing potential using zona-free hamster oocytes.


RESULTS
Percoll -prepared human sperm suspensions exhibited a competence for PMA-induced reactive oxygen species generation which was significantly correlated with leukocyte contamination. However, the purified spermatozoa remaining after paramagnetic bead treatment, also demonstrated an intrinsic capacity for PMA-responsive reactive oxygen species generation and, freed from the oxidative stress created by the leukocytes, exhibited a significantly enhanced capacity for sperm-oocyte fusion.


CONCLUSIONS
Although human spermatozoa can generate reactive oxygen species, sperm function is inhibited by the additional oxidative stress created by contaminating leukocytes. Removal of these cells with paramagnetic beads enhances fertilizing potential.
Aitken
R J
RJ
MRC Reproductive Biology Laboratory, Centre for Reproductive Biology, Edinburgh, Scotland.
Buckingham
D W
DW
West
K
K
Brindle
J
J
D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D003102:Colloids; D052203:Ferrosoferric Oxide; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D007501:Iron; D007962:Leukocytes / Q000276:immunology / Q000378:metabolism*; D008297:Male; D008863:Microspheres; D010087:Oxides; D011205:Povidone; D017382:Reactive Oxygen Species / Q000276:immunology / Q000378:metabolism*; D012822:Silicon Dioxide; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism*
D003102:Colloids; D010087:Oxides; D017382:Reactive Oxygen Species; C016039:Percoll; D012822:Silicon Dioxide; D007501:Iron; D011205:Povidone; D052203:Ferrosoferric Oxide