| Cellular and microenvironmental features | 2-dimensional culture | 3-dimensional culture (advantages) |
| Cell morphology | Typically flattened and unconstrained spreading on the substrate | Depending on the adhesion ligands used, cells can be spherical, spindle shaped or exist as multicellular aggregates |
| Cytoskeletal structure | Display thick bundles of well-developed ventral stress fibers | Few thin stress fibers found near the cell cortex |
| Gene and protein expression | Similarities with in vivo counterparts are lacking | Generally more in vivo-like |
| Cell differentiation | Two-dimensional inducts cell differentiation | Three-dimensional space is closer to the intracellular microenvironment |