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AIM:To generate dendritic cells(DCs)from human pe-ripheral blood and to detect the expression of dendriticcell-specific intercellular adhesion molecule 3 grabbingnonintegrin(DC-SIGN;CD209)for the further study ofDC-SIGN in hepatitis C virus(HCV)transmission.METHODS:Peripheral blood monocytes were isolatedfrom blood of healthy individuals by Ficoll—Hypaquesedimentation and cultured in complete medium contain-ing rhGM-CSF and rhIL-4.Cells were cultured for sevendays,with cytokine addition every two days to obtainimmature DCs.Characteristics of the cultured cells wereobserved under light and scanning microscope,and theexpression of DC-SIGN was detected by immunofluores-cence staining.RESULTS:After seven-day culture,a large number ofcells with typical characteristics of DCs appeared.Theircharacteristics were observed under light and scanningelectron microscope.These cells had a variety of cellshapes such as those of bipolar elongate cells,elaboratestellate cells and DCs.DC-SIGN was detected by immu-nofluorescence staining and its expression level on culti-vated dendritic cells was high.CONCLUSION:DCs with a high expression of DC-SIGNcan be generated from human peripheral blood mono-cytes in complete medium containing rhGM-CSF andrhIL-4.
AIM: To generate dendritic cells (DCs) from human pe-ripheral blood and to detect the expression of dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin (DC-SIGN; CD209) for the further study of DC-SIGN in hepatitis C virus transmission. METHODS: Peripheral blood monocytes were isolatedfrom blood of healthy individuals by Ficoll-Hypaquesedimentation and cultured in complete medium contain-ing rhGM-CSF and rhIL-4.Cells were cultured for sevendays, with cytokine addition every two days to obtainimmature DCs.Characteristics of the cultured cells wereobserved under light and scanning microscope, and theexpression of DC-SIGN was detected by immunofluorescence-cence staining .RESULTS: After seven-day culture, a large number of cells with typical characteristics of DCs appeared. Theircharacteristics were observed under light and scanningelectron microscope.These cells have a variety of cellshapes such as those of bipolar elongate cells, elaboratestellate cells and DCs. DC-SIGN was detec ted by immu-nofluorescence staining and its expression level on culti-vated dendritic cells was high. CONCLUSION: DCs with a high expression of DC-SIGNcan be generated from human peripheral blood mono-cytes in complete medium containing rhGM-CSF and rhIL-4.