Enrichment of macrophages in cell suspensions of human intestinal mucosa by elutriation centrifugation.
To obtain macrophage-rich cell suspensions from human intestinal mucosa, lamina propria specimens were dissociated by incubating in EDTA-collagenase-DNAase solutions and further purified by counter-current centrifugation. During enzymatic incubation macrophage dissociation was linear over the first 8-10 h, reaching a maximum concentration of 10% of total cells and then it plateaued. Counter-current centrifugation resulted in a 5-fold enrichment of macrophages to a mean of 50% with an average recovery rate of 84%. Yields exceeded 0.69 X 10(5) macrophages/g mucosa. Greater than 90% of these cells phagocytosed Staphylococcus aureus, and could be maintained in culture for up to 8 days. Electron microscopy showed satisfactory preservation of the ultrastructure of the macrophages, which also seemed functionally intact.
D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002498:Centrifugation / Q000379:methods; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology*; D008214:Lymphocytes / Q000276:immunology; D008264:Macrophages / Q000276:immunology* / Q000648:ultrastructure; D003012:Microbial Collagenase; D008854:Microscopy, Electron; D010587:Phagocytosis
D003012:Microbial Collagenase