Static three-dimensional culture of human hepatocarcinoma cell with microcarriers

Traditional monolayer culture is still the common method used in hepatocarc inoma cell culture in vitro. For lack of the structure similar to that in v ivo, it is disadvantageous in the research of the relation between structur e and function. We established a three-dimensional (3-D) culture model of h uman hepatocarcinoma cell (BEL-7402) in vitro by using microcarrier cytodex -3 in static condition. The results of SEM, TEM, enzyme activity and flow c ytometry indicated that the cells were polygonal-shaped and arranged in mul tilayers. Intercellular space was 0.5-2.0 mum 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 intracel lular ALT, AST and LDH-L activities were higher in 3-D culture than those o f monolayer culture. Compared with monolayer culture, this 3-D culture with the structure similar to trabecular hepatocarcinoma in vivo is much more s uitable for the research of the interactions of cell-cell and cell-microenv ironment, and might be useful in the investigation of the mechanisms of inv asion, metastasis, and multicellular drug resistance of tumor cells.