3D In Vitro Glioma‐Neuron‐Astrocyte Biomimetic Composites Recapitulate Key Molecular Mechanisms Linked to Glioblastoma Multiforme Pathophysiology

Abstract Glioblastoma multiforme is the most devastating brain tumor without cure. Although in vitro and in vivo research on glioblastoma multiforme have demonstrated its complexity, including interactions with brain cells and the tumor microenvironment, 3D models resembling those key features and allowing to study therapeutic interventions of this aggressive tumor are scarce. Here, a 3D glioblastoma model is developed that establishes a tumor microenvironment including a hyaluronic acid‐based hydrogel cross‐linked with laminin, both of which are key components of the brain's extracellular matrix. This hydrogel mimics the mechanical properties of the brain's extracellular matrix at the macroscopic and mesoscopic levels, as evaluated by stiffness, viscosity using rheological and nanoindentation measurements. The ultra‐soft hydrogel with a storage modulus of 100 Pa is reinforced by 3D printed microfiber scaffolds which allow the setup of a multicellular 3D model including primary cortical neurons and astrocytes and glioblastoma cells. Tumor microenvironment interactions are characterized through nanoindentation and confocal shadow imaging with the 3D in vitro disease model resembling in vivo properties of glioblastoma tumor entities characterized by functional interactions with the surrounding astrocytes and neurons and the tumor's hijacking capability using neuronal signaling to promote its own proliferation.