Spontaneous Hole Formation in Cell Monolayers Emerges from Collective Cell Motion

Abstract: Although cell monolayers typically remain confluent, they can spontaneously develop persistent holes as a result of collective cellular motion. Recent studies on MDCK monolayers cultured on soft substrates have revealed that cells can align to create regions of local nematic order, and topological defects that generate localised mechanical stresses which can spontaneously trigger hole formation. To investigate this process, we develop a continuum multi-phase field model that incorporates internal dissipation and active dipolar forces that drive cell shape anisotropy. Our simulations show that reducing substrate friction enhances cell-cell velocity correlations. In this low-friction regime, topological defects give rise to spiral flow patterns that concentrate stress and can trigger hole formation. We further demonstrate that the number and stability of the holes, whether they close or persist, depends on both substrate friction and cellular activity. These findings underscore the critical role of collective cell dynamics in maintaining tissue integrity.