Isolating pure populations of monocytes from the blood of pregnant women: comparison of flotation in iodixanol with elutriation.
Observations that the innate arm of the immune system is upregulated in pregnancy have highlighted the need for methods of isolating pure populations of monocytes for studies into pregnancy and pre-eclampsia without activating them during the isolation process. Density gradient centrifugation using iodixanol is a useful method for isolating relatively pure populations of unactivated monocytes from human blood but has not been validated in pregnant subjects. We compared the ability of monocytes isolated from pregnant women by density gradient centrifugation using iodixanol (n=6) with monocytes isolated by countercurrent centrifugal elutriation (n=6) in terms of their ability to produce interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) under basal conditions and after stimulation with bacterial lipopolysaccharide (LPS). Under basal conditions, monocytes isolated by density gradient centrifugation produced low amounts of IL-6 and MCP-1. Production of IL-6 and MCP-1 after stimulation of the monocytes with LPS was much greater (p<0.01). There was no statistically significant difference between the two methods in terms of stimulated levels of either cytokine.
Nutt
Jeffrey C
JC
Perinatal and Renal Research Groups, Kolling Institute of Medical Research, Royal North Shore Hospital, St Leonards, NSW, Australia. jnutt@med.usyd.edu.au
Willis
Catherine C
CC
Morris
Jonathan M
JM
Gallery
Eileen D M
ED
D015703:Antigens, CD / Q000276:immunology; D015214:Antigens, Differentiation, Myelomonocytic / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D003287:Contrast Media / Q000494:pharmacology*; D005260:Female; D006801:Humans; D009000:Monocytes / Q000187:drug effects / Q000276:immunology; D011247:Pregnancy; D014283:Triiodobenzoic Acids / Q000494:pharmacology*; D000097382:CD68 Molecule
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D003287:Contrast Media; D014283:Triiodobenzoic Acids; C067980:CD68 antigen, human; D000097382:CD68 Molecule; C044834:iodixanol