Modular, Topographically Patterned, Biomimetic Poly(Ethylene Glycol) Hydrogels as Customized Scaffolds for Organoid Culture

Abstract

The recent development of stem cell-derived, organotypic in vitro models, known as organoids, has revolutionized our ability to study important biological processes in vitro. However, their continued development is limited by the failure of the hydrogel matrices in which they are grown to adequately replicate the tissue-specific ECM cues they experience in their native in vivo environment. Here, we present a highly customizable, modular hydrogel scaffold that can incorporate tissue-specific cues from the extracellular matrix. We demonstrate that these scaffolds can be functionalized with a wide variety of cell adhesion molecules, including peptides and full-length proteins, and can support the attachment and growth of intestinal epithelials organoids, a model organoid system. We also found that these scaffolds can be patterned with large, high-aspect ratio topographical features that mimic anatomical structures (such as intestinal villi) found in vivo. Finally, we show that organoids cultured on these hydrogel scaffolds retain their capacity for multi-lineage differentiation and their ability to model enteric infections. Together, these findings are an excellent proof-of-concept that such hydrogel scaffolds can facilitate the development of organoid models of many organ systems and improve our ability to study a variety of important developmental and pathological processes.