| The poor prognosis of the invasive bladder cancer has correlation with micrometastasis of tumor cells. Dendritic cell-based vaccines has an important role in cancer immunotherapy. In this research, we respectively establish DCs vaccines of loaded with whole tumor antigen and spontaneous metastasis model of bladder cancer. Meanwhile, in order to find out a therapy method for the patients with invasive bladder cancer, we try to explore the effects of DCs vaccines on hematogenous micrometastasis of bladder cancer. The study consists of three parts.Part one : Induction and Culturing of Dendritic Cells Derived from Human Peripheral Blood Monocytes in VitroObjective: To generate high purity and maturity DCs from human peripheral blood monocytes in vitro.Methods: PBMCs were isolated directly from whole blood by density gradient centrifugation and purified by collecting the attached cell ; DCs were then generated by induction and culturing PBMCs for five days with RPMI 1640 medium containing rhGM-CSF and rhIL-4 in vitro, and under the condition of 37℃,5%CO2. On the fifth day, rhTNF-ɑwas added into DCs cultures, which were then incubated for three additional days. The morphology was monitored by light microscopy and transmission electronic microcopy, and the phenotypes were determined by FCM.Results: After eight days of culture, the cells developed typical and significant dendritic morphology and plenty of cells expressed CD1a, CD80 and CD83, features of DCs. Including (78.07±9.43)%CD1a, (60.11±20.50)%CD80 and (46.82±14.15)%CD83 were expressed. About (3.12±1.30)×106 DC cells were derived from 40ml human peripheral blood.Conclusion: This way to generate DCs is simple and easy. The DCs produced by this method acquired high purity and maturity antigenic characteristics of DCs.
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