In general, monolayer culture has been the most popular in vitro model in tumor biology research. In order to mimic the hypoxic and acidic tumor microenvironment, we have established a 3D tumor spheroid culture in in 96-well low adhesion cell culture plates as shown in Figure 14. The 3D multicellular spheroids can build metabolic gradients that provide a more physiologically relevant model than that of 2D cell culture.

Figure 14. 3D tumor spheroid culture.