Fresh hepatocarcinoma tissue and spleen samples were taken from patients during surgery. B cells from spleen were purified and activated. The hepatocarcinoma vaccine was made by cell fusion between hepatic tumor cells and activated B cells. PEG was used as the fusion agent. The fusion cells were cultured and deactivated. MHC Ⅱ and B7 molecules on activated B cells were determined by flow cytometry. Fusion rate and recovery rate of cells after refrigeration were determined respectively at the same time. The results showed that MHC Ⅱ and B7 molecules on the activated B cells were enhanced comparing with B cell. The fusion rates of three cases were 66 84%, 74 43%, and 76 55%, respectively. The recovery rates of cells were 95% and 97% after DMSO and glycerol refrigeration, respectively. The results suggest that the hepatocarcinoma vaccine owns high fusion rate and recovery rate of cells after refrigeration. It's easy to make and store. So the hepatocarcinoma vaccine is suitable for clinical use.