In vitro induction and proliferation of bone marrow-derived dendritic cells in the DA rats

Objective To explore methods of inducing bone marrow-derived dendritic cells (DCs) in Dark Agouti (DA) rats and to provide the theory evidence for research of DCs function and application.Methods Bone marrow cells were taken from the hind limbs and co-cultured with cytokine GM-CSF and IL-4. Ten days later, the morphological characters of DC were observed by inversion fluorescence microscope to be treated with LPS for 1～2 days. The expressions of CD80 and CD86 were detected by flow cytometry to determine DC phenotype. Results The cultured bone marrow cells exhibited morphological characteristics of DCs, having big and long endochylema ecphyma with dendrite type. Eleven days after culture, the cells had DC phenotype as characterized by 85.93% and 81.38% positive expression rate of the co-stimulating molecule CD80 and CD86.Conclusions The approaches of culturing bone marrow-derived cells with GM-CSF and IL-4 could obtain adequate DCs with high purity leading to success in vitro induction and proliferation of DCs. This method paves the way for research in function of DCs and its action in transplantation toleration.