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Objective To induce and culture the dendritic cells in the peripheral blood of breast cancer patients and research on their phenotype. Methods Mononuclear cells were isolated by Ficoll Hypaque centrifugation from 32 breast cancer patients peripheral blood. These cells were plated in six well culture plates(10 6/ml,2 ml/well) in RPMI 1640 medium supplemented with 10% heat inactivated fetal bovine serum,100 ng/ml GM CSF,20 ng/ml IL 4,and/or 20 ng/ml TNF α.Two hours later, nonadherent cells were gently removed and fresh medium was added. Cultured cells were analyzed by flow cytometry with fluorescence labeled monoclonal antibodies. Pictures of cultured and fluorescence stained cells were taken by confocal scanning microscope. Results The diameter of the cells was between 10 and 20 micron. Cells displayed a characteristic CD1a +,CD40 +,CD80 +, CD86 + and CD83 + phenotypes. All of these molecules were not specific for dendritic cells. CD1a and CD83 molecules could also be expressed on the surface of CD3 + T lymphocytes and CD19 + B lymphocytes, especially on activated lymphocytes. Conclusion The molecules of CD1a and CD83 are not specific phenotypes for dendritic cells. Currently, we still need to apply both cell morphology and costimulatory molecules such as CD40,CD80, and CD86 to the identification of dendritic cells.
Objective To induce and culture the dendritic cells in the peripheral blood of breast cancer patients and research on their phenotype. Methods Mononuclear cells were isolated by Ficoll Hypaque centrifugation from 32 breast cancer patients’ peripheral blood. These cells were plated in six well culture plates ( 10 6 / ml, 2 ml / well) in RPMI 1640 medium supplemented with 10% heat inactivated fetal bovine serum, 100 ng / ml GM CSF, 20 ng / ml IL4 and / or 20 ng / Later, nonadherent cells were gently removed and fresh medium was added. Cultured cells were analyzed by flow cytometry with fluorescence labeled monoclonal antibodies. Pictures of cultured and fluorescence stained cells were taken by confocal scanning microscope. Results The diameter of the cells was between 10 and 20 micron. Cells displayed a characteristic CD1a +, CD40 +, CD80 +, CD86 + and CD83 + phenotypes. All of these molecules were not specific for dendritic cells. CD1a and CD83 molecules coul d also be expressed on the surface of CD3 + T lymphocytes and CD19 + B lymphocytes, especially on activated lymphocytes. Conclusion Currently, both molecules of CD1a and CD83 are not specific phenotypes for dendritic cells. Currently, we still need to apply both cell morphology and costimulatory molecules such as CD40, CD80, and CD86 to the identification of dendritic cells.