Sperm-macrophage interaction in the mouse: a quantitative assay in vitro using 111indium oxine-labeled sperm.
The role of reproductive tract macrophages in contraception and reproductive failure has become widely recognized. However, in vitro analysis of sperm phagocytosis by macrophages has relied upon a semi-quantitative method of sperm counting that is of limited accuracy and reproducibility. We have developed an assay using murine sperm labeled with 111indium oxine, and results indicate the labeling to be rapid and efficient. Incorporation of 111indium into sperm increased the dose and sperm concentration and reached 90% maximal uptake after 15 min incubation, with maximal uptake occurring at 30 min. No decrease in sperm motility was noted with levels of oxine in excess of those required for significant labeling. Maximal labeling efficiency occurred in phosphate-buffered saline (PBS), with Dulbecco's modified Eagle's medium (DMEM) + 10% adult bovine serum (ABS) producing significantly less uptake. Label dissociation was detectable in PBS at room temperature, but at 37 degrees C in DMEM + 10% ABS, loss of label occurred at a rate of 23.5%/h. Addition of labeled sperm to murine macrophage monolayers under optimal conditions resulted in uptake of 111indium by macrophages, while free label was unincorporated. Results indicated assay specificity for macrophage-limited uptake, with insignificant label uptake by nonphagocytic murine fibroblasts and better sensitivity than sperm counting. Macrophages from Bacillus Calmette-Guerin (BCG)-infected mice resulted in a decrease in sperm uptake. Female macrophages showed greater capacity for sperm uptake than those of the male mouse. These initial studies demonstrated the utility of this model system in enhancing the understanding of sperm-macrophage interaction in the female reproductive tract.
Department of Obstetrics and Gynecology, Duke University Medical Center, Durham, North Carolina 27710.
D L
DL
Olive
J B
JB
Weinberg
A F
AF
Haney
D000818:Animals; D002469:Cell Separation; D005260:Female; D007205:Indium Radioisotopes; D008262:Macrophage Activation; D008264:Macrophages / Q000502:physiology*; D008297:Male; D051379:Mice; D010587:Phagocytosis; D012737:Sex Factors; D013094:Spermatozoa / Q000276:immunology*
D007205:Indium Radioisotopes