In vitro culture and characterization of mouse spleen dendritic cells

    TIAN Rong, LI Wei, YU Jiª²Yun, LIU Yuª²Feng

    Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi¬ðan 710033, China, Institute of Basic Medicine, Military Academy of Medical Sciences, Beijing 100850, China

    ¡¾Abstract¡¿ AIM: To explore and optimize the methods for in vitro culture of mouse dendritic cells (DC) that were then observed morphologically and identified biologically. METHODS: Mice were injected with 200 mg/kg cyclophosphamide through the tail vein, and were sacrificed 8 d later. Spleen cells were cultured with ordinary methods firstly, and 3 d later conditional medium containing ILª²4, GMª²CSF was added, and TNFª²¦Á was added at day 5. At day 8, cells were subjected to phaseª²contrast microscope, scanning electron microscope, and transmission electron microscope analysis. At day 3, 5, 7, cells were subjected to FACS for detection of cell surface markers. RESULTS: Cultured cells displayed a typical DC phenotype in morphological analysis, and FACS showed these cells expressing such DC markers as CD11c, CD86,¢ñAb, H2D6. CONCLUSION: Using of ILª²4, GMª²CSF, TNFª²¦Á as stimulators in the mouse spleen cell culture can obtain DC with high quality and high purity, which makes a foundation for future research on the basic and clinical usage of DC. ......