Study on Isolation and Biology of Rat Bone Marrow-Derived Dendritic Cell in vitro

Objective To explore methods for isolation of rat bone marrow-derived dendritic cell(DC) and to investigate its bionomics.Methods Bone marrow cells were taken from the hind limb and co-cultivated with cytokine GM-CSF and IL-4 for 5 days.The morphological characters of DC were observed by a inversed microscope,the cells were stimulated for 48 hours then morphological characters were observed again.Its MHC-Ⅱ and CD80 molecules were detected with FCM,its IL-6 and IL-12 in co-culture medium was quantified by ELISA.Results Cultivated DC derived from bone-marrow cells have ecphyma and morphological characteristics of DC.MHC-Ⅱ and CD80 phenotype were characterized by 57.3% and 29.1% positive expression rate on untreated DC 7 days later,and 70.9% and 73.5% on DC treated with LPS.The quantity of IL-6 and IL-12 in LPS stimulated culture medium was higher than unstimulated(P0.01).Conclusion A approach of DC derived from bone-marrow cell is established,DC change phenotype and secrete cytokine are discovered maturation in vitro.