Purification of human monocytes by adherence to polymeric fluorocarbon. Characterization of the monocyte-enriched cell fraction.
Human mononuclear cells were obtained from peripheral blood by density gradients. Monocytes can be purified after cultivation of 2 hours by a modified adherence procedure on membranes of gas-permeable polymeric fluorocarbon (teflon). After further cultivation of 24-48 hours, monocyte-enriched cell fraction can be easily detached from the membranes with a viability greater than 98% and a final cell yield of approximately 50% of the peripheral monocyte count. The cells showed the morphological and cytochemical characteristics of human monocytes and differentiated into dense monolayers of macrophages within 10 days of cultivation. Immune-autoradiography with iodine-125-labelled xenogeneic antimonocytic antisera and staining with several monoclonal antisera in an indirect immunofluorescence technique revealed up to 92% of these cells to carry monocytic characteristics. To show their functional integrity, monocytes obtained by this procedure were activated by 48 hours' cultivation with synthetic alkyl-lysophospholipids to inhibit the proliferation of autologous tumor cells.
D001345:Autoradiography; D001402:B-Lymphocytes / Q000276:immunology; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D008246:Lysophospholipids; D009000:Monocytes / Q000032:analysis / Q000276:immunology; D010587:Phagocytosis; D010743:Phospholipids / Q000494:pharmacology; D011138:Polytetrafluoroethylene / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000276:immunology
D008246:Lysophospholipids; D010743:Phospholipids; D011138:Polytetrafluoroethylene