Static three-dimensional culture of human hepatocarcinoma cell with microcarriers

Traditional monolayer culture is still the common method used in hepatocarcinoma cell culturein vitro. For lack of the structure similar to thatin vivo, it is disadvantageous in the research of the relation between structure and function. We established a three-dimensional (3-D) culture model of human hepatocarcinoma cell (BEL-7402)in vitro by using microcarrier cytodex-3 in static condition. The results of SEM, TEM, enzyme activity and flow cytometry indicated that the cells were polygonal-shaped and arranged in multilayers. Intercellular space was 0.5–2.0 μm wide where lots of microvilli could be seen. Adjacent cells were connected with desmosomes and localized membrane projects. More than 90% of the cells were viable and maintained the consumption of glucose and the expression of EGF receptor. The intracellular ALT, AST and LDH-L activities were higher in 3-D culture than those of monolayer culture. Compared with monolayer culture, this 3-D culture with the structure similar to trabecular hepatocarcinomain vivo is much more suitable for the research of the interactions of cell-cell and cell-microenvironment, and might be useful in the investigation of the mechanisms of invasion, metastasis, and multicellular drug resistance of tumor cells.