Induction and identification of rabbit peripheral blood derived dendritic cells

Jing Zhou; FuYuan Yang; WenLi Chen

doi:10.1117/12.907064

28 February 2012 Induction and identification of rabbit peripheral blood derived dendritic cells

Jing Zhou, FuYuan Yang, WenLi Chen

Proceedings Volume 8214, Advanced Biomedical and Clinical Diagnostic Systems X; 82141A (2012) https://doi.org/10.1117/12.907064
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

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.

Citation Download Citation

Jing Zhou, FuYuan Yang, and WenLi Chen "Induction and identification of rabbit peripheral blood derived dendritic cells", Proc. SPIE 8214, Advanced Biomedical and Clinical Diagnostic Systems X, 82141A (28 February 2012); https://doi.org/10.1117/12.907064

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