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Despite the importance of trophoblast invasion for processes essential at early stages of pregnancy, much remains unknown regarding the balance of signaling molecules that may influence trophoblast invasion into the endometrium. The goal of this study was to use three-dimensional trophoblast spheroid motility assays to examine the effect of cues from the maternal\u2013fetal interface on trophoblast motility. We report use of a methacrylamide-functionalized gelatin hydrogel to support quantitative analysis of trophoblast outgrowth area and cell viability. We show that this multidimensional model of trophoblast motility can resolve quantifiable differences in outgrowth area and viability in the presence of a known invasion promoter, epidermal growth factor, and a known invasion inhibitor, transforming growth factor \u03b21. We then investigate the sensitivity of trophoblast motility to cortisol, a hormone associated with exogenous stressors. Together, this approach provides a toolset to investigate the coordinated action of physiological and pathophysiological processes on early stages of trophoblast invasion.<\/jats:p>","DOI":"10.1089\/ten.tea.2020.0097","type":"journal-article","created":{"date-parts":[[2020,11,20]],"date-time":"2020-11-20T16:52:46Z","timestamp":1605891166000},"page":"1064-1073","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":20,"title":["Tuning Trophoblast Motility in a Gelatin Hydrogel via Soluble Cues from the Maternal\u2013Fetal Interface"],"prefix":"10.1177","volume":"27","author":[{"given":"Samantha G.","family":"Zambuto","sequence":"first","affiliation":[{"name":"Department of Bioengineering, University of Illinois Urbana-Champaign, Urbana, Illinois, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathryn B.H.","family":"Clancy","sequence":"additional","affiliation":[{"name":"Department of Anthropology, University of Illinois Urbana-Champaign, Urbana, Illinois, USA."},{"name":"Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign, Urbana, Illinois, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brendan A.C.","family":"Harley","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering, University of Illinois Urbana-Champaign, Urbana, Illinois, USA."},{"name":"Carl R. 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