Previously a cell line MHCC97H with high metastatic potential was established for HCC metastatic study. For further analysis of the biological character and the metastatic potential of HCC cells in 3D state, a HCC spheroid with tissue characteristics derived from MHCC97H cells formed in a RWV bioreactor system with PLGA scaffold. The spheroid was transparent and compact in macroscopical morphology, and the cells of it displayed concentric arrangement of large proliferating tumor cells in the periphery and smaller non-proliferating cells in deeper regions, as well as some cells necrosis in central section. The spheroid also had a tissue-like ultra-structure desmosome junction and tight junction complex at membrane borderline of adjacent cells. The higher contents of ALB, AFP, γ-GT in culture supernatant of HCC spheroid implied that the spheroid better regained the function of HCC specific proteins/enzymes synthesis and secretion. Moreover, the similar changes of ALB, AFP, γ-GT in culture medium were frequently found in serum contents of HCC patients in clinic. The expression patterns of adhesion molecules, HCC specific gene and metastasis associated genes in the established spheroid clearly demonstrated that the spheroid was not only a HCC model, but also a model with high metastasis potential. More significantly, the occurrence of secondary tumor nodule around primary tumor in liver and the tumor cells of metastatic lesions in lungs of the tested nude mice verified intrahepatic metastasis and distant metastasis. All the results indicated that the establishment of a HCC tissue-like spheroid model was successful and it might have applications in HCC study, anticancer drug screening and establishment of animal model.