Surface modification plays an essential role in dictating cell behavior and fate, as it creates a microenvironment that profoundly influences cell attachment, migration, proliferation, and differentiation. This review aims to the intricate interplay of culture surface properties, including topography, stiffness, charge, and chemical modifications, demonstrating their profound impact on cell destiny. We explore the nuanced responses of cells to varying surface topographies, from nano- to microscale features, elucidating the influence of geometric patterns and roughness. We also investigate the impact of substrate stiffness, highlighting the way cells perceive and respond to mechanical cues mimicking their native environments. The role of surface charge is examined, revealing how electrostatic interactions influence cell adhesion, signaling, and cell fate decisions. Finally, we delve into the diverse effects of chemical modifications, including the presentation of bioactive molecules, growth factors, and extracellular matrix (ECM) components, demonstrating their ability to guide cell behavior and stimulate specific cellular responses. This review offers comprehensive insights into the important role of surface properties in shaping cell fate, offering promising avenues for developing sophisticated cell culture platforms for applications in drug discovery, regenerative medicine, and fundamental research.