Two-parameter contributionso fs tiffness and wettability display alteredb ehavior of hBM-MSCs as compared to solely investigating the wettability of the biointerface with ah omogenous stiffness. The importance of double parameter influence is identified by using newly developed double linear wettability-stiffness gradients. The interfaces show that substrate stiffness can influence cell adhesion and spreading both positivelya nd negatively in combination with wettability.T his effect is validated by decoupling the stiffness gradientu sing aw ettability gradient composed of amonolayer on glass ensuring ahomogenous stiffness.

New high‐throughput technologies for cell–material interaction studies provide researchers with powerful tools to speed up research in the field of biomaterial–cell interactions. However, sharing technologies is often difficult due to the necessity of specific knowledge and experiences. Engineered surfaces can elucidate effects of surface topography on cell behavior, which is of critical value for gaining control over cellular processes. Here, the translation of a gradient‐based high‐throughput cell screening approach for aligned nano/micro topographies interacting with cells is presented. An aligned topography 96‐well plate is created by upscaling of highly specific gradient technology. The resulting cell culture dishes are compatible with general laboratory and imaging equipment, and the platform allows for studying cell behavior with regard to adhesion and alignment. The challenge lies in increasing the dimensions of the previous 1 × 1 cm gradient topography substrate, to be able to cover the span of a 96‐well plate and translate it into a standardized cell‐screening tool. Adhesion experiments of human bone marrow derived mesenchymal stem cells confirm the standardization, compatibility, and usability of the technology. In the process of using multi‐system imaging and analysis, it becomes apparent that future challenges need to include universally applied data analysis approaches.