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Updated: Aug 10, 2026
Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
Published on: October 18, 2016
Expression of dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin on dendritic cells
Jun Li1, Zhi-Hua Feng, Guang-Yu Li
1The Center of Diagnosis and Treatment for Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, Shaanxi Province, China.
Insights
Human peripheral blood monocytes can be cultured to generate dendritic cells (DCs) that highly express dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin (DC-SIGN). This method aids in studying DC-SIGN
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) play a crucial role in immune responses.
- Dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin (DC-SIGN) is implicated in pathogen transmission, including Hepatitis C Virus (HCV).
Purpose of the Study:
- To establish a method for generating dendritic cells (DCs) from human peripheral blood.
- To detect the expression of DC-SIGN on these generated DCs for further research into its role in HCV transmission.
Main Methods:
- Isolation of peripheral blood monocytes from healthy donors.
- Culture of monocytes in complete medium with rhGM-CSF and rhIL-4 for seven days to generate immature DCs.
- Observation of cell morphology via light and scanning electron microscopy.
- Detection of DC-SIGN expression using immunofluorescence staining.
Main Results:
- Successful generation of a large number of cells exhibiting typical dendritic cell characteristics after seven days of culture.
- Observation of diverse cell morphologies, including bipolar, stellate, and typical DC shapes.
- High expression levels of DC-SIGN detected on the generated dendritic cells via immunofluorescence.
Conclusions:
- Dendritic cells (DCs) expressing high levels of DC-SIGN can be effectively generated from human peripheral blood monocytes.
- The described culture method using rhGM-CSF and rhIL-4 provides a reliable source of DCs for studying DC-SIGN function in contexts like HCV transmission.
Aim:
To generate dendritic cells (DCs) from human peripheral 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 (HCV) transmission.
Methods:
Peripheral blood monocytes were isolated from blood of healthy individuals by Ficoll-Hypaque sedimentation and cultured in complete medium containing rhGM-CSF and rhIL-4. Cells were cultured for seven days, with cytokine addition every two days to obtain immature DCs. Characteristics of the cultured cells were observed under light and scanning microscope, and the expression of DC-SIGN was detected by immunofluorescence staining.
Results:
After seven-day culture, a large number of cells with typical characteristics of DCs appeared. Their characteristics were observed under light and scanning electron microscope. These cells had a variety of cell shapes such as those of bipolar elongate cells, elaborate stellate cells and DCs. DC-SIGN was detected by immunofluorescence staining and its expression level on cultivated dendritic cells was high.
Conclusion:
DCs with a high expression of DC-SIGN can be generated from human peripheral blood monocytes in complete medium containing rhGM-CSF and rhIL-4.
