In the analyzed volume, regions spaced below 20 nm, within molecular range, represented 31% of the macrophage membrane surface and more than 27% of the myotube membrane. This revealed a new type of interaction between macrophages and myogenic cells by direct heterocellular surface apposition over large areas and long linear distances. Ultrastructural analysis was performed by conventional transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) followed by 3D modeling of electron tomography (ET) data. Samples harvested (3 and 5 days) post-injury were screened by light and confocal microscopy. We tested this hypothesis during different steps of skeletal muscle regeneration: (a) the recruitment of activated SC (b) the differentiation of MPC (c) myotubes growth, in a mouse model of crush injury. Our hypothesis was that a direct and close cellular interaction between SC/MPC and invading myeloid cells is a key step to control regeneration. Furthermore, intercellular connections during skeletal muscle regeneration have not been previously thoroughly documented. However, the regulatory mechanism during this process is still under evaluation, with the final aim to manipulate regeneration when the intrinsic mechanism is corrupted. Regeneration in adult skeletal muscle relies on the activation, proliferation, and fusion of myogenic precursor cells (MPC), mostly resident satellite cells (SC).
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