The influence of culture conditions and serum lipids on interleukin-1 production by human monocytes.
Reliable assessment of IL-1 production by human monocytes is critically dependent on the methods for isolation and culture of these cells. In the present study, the quality of pipettes and the preparation of Ficoll-Isopaque appear to be crucial for IL-1 production from both LPS-stimulated and unstimulated monocytes. Different brands and lots of polystyrene culture wells give rise to great variation in IL-1 production. When carefully prepared, hydrophobic teflon membranes, to which mononuclear phagocytes poorly adhere, are used as the culture substrate, stimulation of IL-1 production is observed. The HLA DR3 haplotype of the monocyte donors did not influence IL-1 production. The addition of normal human AB serum to the cultures usually increases IL-1 production, although strong inhibition of both unstimulated and LPS-stimulated IL-1 production was also observed after addition of a diet-induced hyperlipemic AB serum. This inhibition was not due to cholesterol, chylomicrons, high- or low-density lipoproteins. When monocytes were cultured at different temperatures, the only abnormality found was a decrease of cell-associated IL-1 at 41 degrees C.
Van der Meer
J W
JW
Department of Infectious Diseases, University Hospital Leiden, The Netherlands.
Van de Gevel
J S
JS
Van Hinsbergh
V W
VW
Leijh
P C
PC
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003470:Culture Media; D006684:HLA-DR Antigens; D006801:Humans; D006949:Hyperlipidemias / Q000097:blood; D007375:Interleukin-1 / Q000096:biosynthesis*; D008055:Lipids / Q000097:blood*; D008264:Macrophages / Q000378:metabolism; D008297:Male; D051379:Mice; D009000:Monocytes / Q000145:classification / Q000378:metabolism*; D010641:Phenotype; D013696:Temperature
D003470:Culture Media; D006684:HLA-DR Antigens; D007375:Interleukin-1; D008055:Lipids