Patkar, Sai S.Garcia, Cristobal GarciaPalmese, Luisa L.Kiick, Kristi L.2023-11-292023-11-292023-08-14Patkar, Sai S., Cristobal Garcia Garcia, Luisa L. Palmese, and Kristi L. Kiick. “Sequence-Encoded Differences in Phase Separation Enable Formation of Resilin-like Polypeptide-Based Microstructured Hydrogels.” Biomacromolecules 24, no. 8 (August 14, 2023): 3729–41. https://doi.org/10.1021/acs.biomac.3c00418.1526-4602https://udspace.udel.edu/handle/19716/33637This document is the Accepted Manuscript version of a Published Work that appeared in final form in Biomacromolecules, copyright © 2023 American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.biomac.3c00418. This article will be embargoed until 08/17/2024.Microstructured hydrogels are promising platforms to mimic structural and compositional heterogeneities of the native extracellular matrix (ECM). The current state-of-the-art soft matter patterning techniques for generating ECM mimics can be limited owing to their reliance on specialized equipment and multiple time- and energy-intensive steps. Here, a photocross-linking methodology that traps various morphologies of phase-separated multicomponent formulations of compositionally distinct resilin-like polypeptides (RLPs) is reported. Turbidimetry and quantitative 1H NMR spectroscopy were utilized to investigate the sequence-dependent liquid–liquid phase separation of multicomponent solutions of RLPs. Differences between the intermolecular interactions of two different photocross-linkable RLPs and a phase-separating templating RLP were exploited for producing microstructured hydrogels with tunable control over pore diameters (ranging from 1.5 to 150 μm) and shear storage moduli (ranging from 0.2 to 5 kPa). The culture of human mesenchymal stem cells demonstrated high viability and attachment on microstructured hydrogels, suggesting their potential for developing customizable platforms for regenerative medicine applications.en-UScellshydrogelsmonomerspeptides and proteinsphase separationArticle