Liyao Xu, Frank Daniel Peters, Haokang Zhang, Madison Stiefbold, Thangam Ramar, Aditya Agarwal, Leo Q Wan
Substrate topography, such as curvature, has been recognized as a key physical cue that regulates cell organization and tissue function, yet how curvature influences chiral cell behavior remains unclear. Here, we created curved surfaces (concave or convex) separated by flat regions to investigate the role of cell chirality in curvature-guided multicellular morphogenesis. C2C12 cells showed opposite alignment biases on concave and convex surfaces, and the cells on flat regions adopted a reversed bias compared to those on the adjacent curved areas. Cells with opposite intrinsic or pharmacologically altered chirality exhibited correspondingly reversed alignment patterns. Detailed analyses revealed that local curvature initially directed cell elongation along distinct geometric axes on convex and concave surfaces, followed by biased tilting and migration governed by intrinsic cellular chirality. Together, our findings demonstrate that cell chirality superimposes a directional bias upon curvature-guided elongation, thereby driving collective alignment and migration. This work provides new insights into how cells respond to curvature and highlights a potential mechanism for tissue-level symmetry breaking, regulated by substrate topography.