Biocompatibility of Hydrogels for Glomerular 3D Co-Culture: A Comparative Analysis

Conventional 2D mono-cultures fall short in replicating the complex microenvironment of glomerular tissue, where cell-cell and cell-matrix interactions are critical. To better mimic in vivo conditions, the development of robust 3D co-culture systems is essential. Here, we systematically evaluate five hydrogel matrices-Matrigel, alginate dialdehyde-gelatin (ADA-GEL), fibrin, recombinantly produced spider silk protein eADF4(C16)-RGD, and allyl-modified gelatin (GelAGE)-for their suitability in supporting glomerular 3D co-culture. The hydrogels are assessed for handling properties, cell viability, and the support of physiological cell behavior using bright-field microscopy, live/dead assays, immunofluorescence, and multiphoton imaging. Among the tested hydrogels, GelAGE and eADF4(C16)-RGD demonstrate superior biocompatibility and structural support. Due to its ease of use and comparable biological performance, GelAGE and spider silk protein eADF(C16)-RGD are selected for further mechanical characterization, revealing favorable viscoelastic properties. These findings position both hydrogels as a promising candidate for engineering physiologically relevant 3D glomerular models and advancing kidney tissue research.