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The rarity of such twinning events, combined with ethical constraints in human embryo research, makes investigating the mechanisms behind twinning practically infeasible. As a result, there is a significant knowledge gap regarding the origins and early phenotypic presentation of monochorionic twin embryos. In this study, a microthermoformed\u2010based microwell screening platform is used to identify conditions that efficiently induce monochorionic twins in human stem cell\u2010based blastocyst models, termed \u201ctwin blastoids\u201d. These twin blastoids contain a cystic GATA3+ trophectoderm\u2010like epithelium encasing two distinct inner cell masses (ICMs). Morphological and morphokinetic analyses reveal that twinning occurs during the cavitation phase via splitting of the OCT4+ pluripotent core. Notably, each ICM in twin blastoids contains its own NR2F2+ polar trophectoderm\u2010like region, ready for implantation. This is functionally tested in a microfluidic chip\u2010based implantation assay with epithelial endometrium cells. Under defined flow regimes, twin blastoids show increased adhesion capacity compared to singleton blastoids, suggestive of increased implantation potential. In conclusion, the development of technology enabling large\u2010scale formation of twin blastoids, coupled with high\u2010sensitivity readout capabilities, presents an unprecedented opportunity for systematically exploring monochorionic twin formation and its impact on embryonic development.<\/jats:p>","DOI":"10.1002\/adma.202313306","type":"journal-article","created":{"date-parts":[[2024,4,9]],"date-time":"2024-04-09T15:29:26Z","timestamp":1712676566000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Monochorionic Twinning in Bioengineered Human Embryo Models"],"prefix":"10.1002","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8670-1470","authenticated-orcid":false,"given":"Dorian G.","family":"Luijkx","sequence":"first","affiliation":[{"name":"MERLN Institute of Technology\u2010Inspired Regenerative Medicine Department for Instructive Biomaterials Engineering (IBE) Maastricht University  Universiteitssingel 40 Maastricht 6229ET The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asli","family":"Ak","sequence":"additional","affiliation":[{"name":"MERLN Institute of Technology\u2010Inspired Regenerative Medicine Department for Instructive Biomaterials Engineering (IBE) Maastricht University  Universiteitssingel 40 Maastricht 6229ET The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ge","family":"Guo","sequence":"additional","affiliation":[{"name":"Living Systems Institute University of Exeter  Exeter EX4 4QD UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clemens A.","family":"van Blitterswijk","sequence":"additional","affiliation":[{"name":"MERLN Institute of Technology\u2010Inspired Regenerative Medicine Department for Instructive Biomaterials Engineering (IBE) Maastricht University  Universiteitssingel 40 Maastricht 6229ET The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1354-0167","authenticated-orcid":false,"given":"Stefan","family":"Giselbrecht","sequence":"additional","affiliation":[{"name":"MERLN Institute of Technology\u2010Inspired Regenerative Medicine Department for Instructive Biomaterials Engineering (IBE) Maastricht University  Universiteitssingel 40 Maastricht 6229ET The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0702-5609","authenticated-orcid":false,"given":"Erik J.","family":"Vrij","sequence":"additional","affiliation":[{"name":"MERLN Institute of Technology\u2010Inspired Regenerative Medicine Department for Instructive Biomaterials Engineering (IBE) Maastricht University  Universiteitssingel 40 Maastricht 6229ET The Netherlands"},{"name":"Gynaecology Women Mother Child Centre Maastricht University Medical Centre+ (MUMC+)  P. 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