Traditional compound effect assays typically involve cells adhered to a plate surface in 2D culture. In this setup, all cells are functionally similar and receive uniform exposure. However, this approach lacks translational relevance since cells naturally grow in 3D structures, resulting in functional diversity and uneven exposure due to factors like oxygen and nutrient gradients. Certain cell lines can form 3Dstructures in vitro under specific conditions, leading to varying growth rates and exposure levels based on location within the structure. Utilizing 3D models enhances translational relevance, enabling assessment of compound effects on proliferation and cytotoxicity in a more physiologically relevant environment