Mechanical stretch induced collective cell alignment

Abstract

Cell alignment is a fundamental process in tissue morphogenesis. While density-dependent collective cell alignment has been widely observed, its underlying mechanisms remain poorly understood. Here, using C2C12 myoblasts, we demonstrate that static uniaxial mechanical stretch induces collective cell alignment in a density-dependent manner: densely populated cultures align robustly, whereas sparse populations do not. We reveal a biphasic alignment process, comprising an initial passive phase and a subsequent active phase. The passive phase, driven by substrate deformation, transiently biases cell orientation along the stretch axis regardless of density. In the active phase, initial alignment progressively dissipates in low-density cultures, but is sustained and reinforced in high-density cultures. Supported by coarse-grained agent-based simulations, we propose that self-generated cellular forces facilitate kinetic transitions between orientations, enabling cells to explore orientational states, whereas cell-cell interactions provide a thermodynamic bias that stabilizes the locally aligned state. In dense cultures, strong intercellular interactions promote this stabilization, enabling persistent alignment. In contrast, sparse cultures lack sufficient cell-cell interaction, leading to alignment dissipation. Within this C2C12 system, our findings highlight the cooperative roles of cellular forces and intercellular interactions in orchestrating multicellular ordering, offering new insights into mechanobiology of tissue morphogenesis.