Proceedings Article | 28 February 2012
Proc. SPIE. 8214, Advanced Biomedical and Clinical Diagnostic Systems X
KEYWORDS: Confocal microscopy, Microscopes, Optical filters, Biomedical optics, Blood, Luminescence, Life sciences, Flow cytometry, Laser scanners, Fetus
Purpose: To study a method of the induction of dendritic cells (DCs) from rabbit peripheral blood. Methods: Peripheral
blood cells were removed from rabbit, filtered through nylon mesh. Peripheral blood mononuclear cells (PBMC) were
isolated from the blood cells by Ficoll-Hypaque centrifugation (density of 1.077g/cm3).To obtain DCs, PBMC were
cultured in RPMI1640 medium containing 10% fetal calf serum, 50U/mL penicillin and streptomycin, referred to
subsequently as complete medium, at 37°C in 5% CO2 atmosphere for 4 hours. Nonadherent cells were aspirated,
adherent cells were continued incubated in complete medium, supplemented with granulocyte/macrophage
colony-stimulating factor (GM-CSF, 50ng/ml),and interleukin 4 (IL-4, 50ng/ml) for 9 days. Fluorescein labeled
antibodies(anti-CD14, anti-HLA-DR, anti-CD86) were used to sign cells cultured for 3,6,9 days respectively, Then flow
cytometry was performed. Results: Ratio of anti-HLA-DR and anti-CD86 labeled cells increased with induction time
extension, in contrast with anti-CD14. Conclusion: Dendritic cells can be effectively induced by the method of this
experiment, cell maturation status increased with induction time extension.
