Murine dendritic cells in vitro and identification of.doc

 PAGE \* MERGEFORMAT 14 Murine dendritic cells in vitro and identification of Authors: Rong Tian, Li Wei, Yu Ji-yun, LIU Yu-feng [Keywords:] dendritic cells In vitro culture and characterization of mouse spleen dendritic cells [Abstract] 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 IL4, GMCSF was added, and TNFα was added at day 5. At day 8, cells were subjected to phasecontrast 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, Ⅰ Ab, H2D6. CONCLUSION: Using of IL4, GMCSF, TNFα 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. [Keywords] dendritic cells; cytokines; MHC Ⅱ molecule [Abstract] Objective: To explore and optimize the in vitro induction and expansion of mouse dendritic cells (DC) method, and were observed and biological identification. Methods: cyclophosphamide 200 mg / kg via the tail vein injection of Balb / c mice, 8 d after the mice were killed, spleen cells of conventional culture, the first 3 to join containing IL4, GMCSF cytokine conditioned medium, the 5th day of additional TNFα , No. 8, preparation of specimens for phase-contrast microscopy, scanning electron microscope , transmission electron microscopy observation, respectively, in developing the first 3, 5, 7, mature cells by flow cytometry surface ma