The effect of macromolecular crowders on deposition of extracellular matrix in astrocyte cultures

Macromolecular crowding (MMC) is a biophysical phenomenon that has proven effective in enhancing extracellular matrix (ECM) deposition in vitro. However, MMCs potential in neuroglial cell cultures remains underexplored. This study investigates the effects of three distinct MMC agents [carrageenan (CR), dextran sulphate (DxS) and FicollⓇ cocktail (FC)] on ECM deposition and cell behaviour of Neu7 and primary astrocytes. While the viability and metabolic activity of Neu7 astrocytes were unaffected by any of the crowding agents, primary astrocytes exhibited a significant decrease in viability and metabolic activity in the presence of CR and DxS. The addition of CR, DxS, and FC resulted in a significant increase in deposition of fibronectin, collagen IV, collagen I, GFAP and CS56 in Neu7 astrocytes. In primary astrocytes, FC significantly enhanced the expression of astrocytic markers and increased the deposition of ECM proteins, including fibronectin and collagen IV. This study highlights the advantages of using FC as a MMC agent for enhancing ECM deposition in astrocytes. The method demonstrates potential for developing fast and more physiologically relevant in vitro models and improving drug screening processes for future studies. The observed benefits underscore the utility of FC in creating advanced cellular models that better mimic the native neural environment.