Spheroid Formation Protocol

In vitro 3D cell culture models are widely recognized as more physiologically relevant systems compared to 2D formats. To recapitulate features of native tumor microenvironments, cancer cells can be cultured in Corning spheroid microplates, which combine the Corning Ultra-Low Attachment surface with innovative well geometry to provide an ideal tool for generating, culturing, and assaying 3D multicellular spheroids in the same plate, without the need for a transfer step. This protocol describes a basic method for generating and culturing tumor spheroids in a 96-well spheroid microplate format. This basic protocol for culture and assay can be adapted for all spheroid microplates.