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Updated: Aug 24, 2026
Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
Published on: October 18, 2016
[The effect of calcium ionophore on dendritic cells derived from peripheral blood mononuclear cells]
Jun Wu1, Xiao-huai Wang, Jie Wang
1Institute of Molecular Oncology, General Hospital of Guangzhou Command, Guangzhou 510010, China. Junwuwang@hotmail.com
Aim:
To explore the effect of calcium ionophore (CI) on dendritic cells (DC) derived from peripheral blood monocytes.
Methods:
Peripheral blood monocytes from healthy donors were treated with 100 microg/L rhGM-CSF, 100 microg/L rhGM-CSF plus (10 microg/L) CI, and 100 microg/L rhGM-CSF plus (100 microg/L) respectively. After cultivation of 40 hours, cellular morphology was observed under light microscope and electron microscope. Surface markers on treated PBMCs were analyzed by flow cytometry. MTT colorimetry was used to detect proliferation of autologous T cells.
Results:
Peripheral blood monocytes treated with 100 microg/L rhGM-CSF plus 100 microg/L CI for 40 hours showed typical morphology of DCs. Simultaneously there was a decrease in CD14 expression and increase in HLA-DR, CD40, CD83 and CD86 expressions on these cells. In addition, PBMCs treated with 100 microg/L rhGM-CSF pass 100 microg/L CI for 40 hrs. Could evidently stimulate proliferation of autologous T cells.
Conclusion:
CI can markedly enhance transformation of peripheral blood monocytes induced by GM-CSF into DCs.
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