Validation of a Novel Tumoroid-Based Cell Culture Model to Perform 3D in vitro Cell Signaling Analyses

Here we demonstrate an in vitro HTRF® microplate assay that can quantify total, as well as phosphorylated elF4E. The assay used a novel 3D culture system called RAFT' to create cell accumulations termed tumoroids in a cell/collagen hydrogel mix. In addition, the assay was also performed with cells cultured using traditional 2D methods for comparison. Dispensing and removal steps were performed by the MultiFloTM FX Microplate Dispenser, including cell/collagen mix, medium, and reagent dispensing, as well as removal of spent medium and compounds. Detection of the fluorescent signals from the HTRF assay, as well as tumoroid imaging, was performed by the CytationTm 3 Cell Imaging Multi-Mode Reader. Validation data generated using RAFT confirms the robustness of the 3D system. Pharmacology comparison data from both methods also demonstrates the validity of using 3D cell culture for cell signaling analyses.