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To evaluate whether reduced permeability was a driving factor in reduced cell viability in synthetic hydrogels, we compared poly(ethylene glycol) vinyl sulfone (PEG\u2010VS) hydrogels with Matrigel to quantify the influences of modulus, transport, and confinement on encapsulated cells. We observed extensive reductions in cell viability when encapsulated in PEG\u2010VS gels compared to Matrigel. In transwell experiments that decouple hydrogel\u2010restricted serum from cell\u2010gel adhesion, serum restriction reduced cell viability, matching the cell viability observed in 3D cultures. Our unique combination of 2D and 3D hydrogel\u2010based cell cultures provides a framework for investigating the intersecting effects of the cell microenvironment's properties on cell viability. This work demonstrates that biomaterial\u2010restricted protein transport is a critical design consideration when using synthetic 3D cell culture hydrogels.<\/jats:p>","DOI":"10.1002\/adma.202521245","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T15:11:07Z","timestamp":1772982667000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The Intersecting Physical Mechanisms That Regulate Cell Viability in 3D Synthetic Hydrogels"],"prefix":"10.1002","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5091-3321","authenticated-orcid":false,"given":"Nathan R.","family":"Richbourg","sequence":"first","affiliation":[{"name":"The University of Massachusetts at Amherst  Amherst Massachusetts USA"},{"name":"Tufts University  Medford Massachusetts USA"},{"name":"FAMU\u2010FSU College of Engineering  Tallahassee Florida USA"},{"name":"Florida State University  Tallahassee Florida 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