ECM stiffness is a potential instructive cue during capillary morphogenesis. Bulk measurements have often been used to investigate matrix stiffness as a cellular cue. However, local peri-cellular stiffness can be heterogeneous and vary greatly from measured bulk properties spatially and temporally. Here, we utilize bulk rheology and optical tweezers active microrheology (AMR) to investigate the dynamic mechanical crosstalk between cells and the surrounding matrix during capillary morphogenesis across length scales with various supporting stromal cells. We will present our current findings focusing on the discrepancies between bulk and microrheological distributions of measured stiffness across different stromal cell types.
Studies of cell-extracellular matrix (ECM) interactions within fibrous systems such as collagen or fibrin are challenging, particularly if peri-cellular stiffness cannot be monitored. Here we present our light-based method for non-invasive patterning of molecular crosslinking combined with multi-axes optical tweezers active microrheology to map ECM stiffness landscapes. This method allows us to generate prescribed stiffness gradients and associated anisotropies, which model stiffness of the natural peri-cellular ECM. Patterned crosslinking induces strain hardening and measured stiffness gradients are in agreement with predicted strain fields. Migratory cells respond to these gradients as assessed by change in F-actin distribution and morphological properties.
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