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Also, pre-optimization of single spheroid properties is often neglected. Here, we report a simple setup for rapid biomanufacturing of spheroid-based patch-like vascular beds. Prior to patch assembly, spheroids combining mesenchymal stem\/stromal cells (MSCs) and outgrowth endothelial cells (OECs) at different ratios (10:1; 5:1; 1:1; 1:5) were formed in non-adhesive microwells and monitored along 7 d. Optimal OEC retention and organization was observed at 1:1 MSC\/OEC ratio. Dynamic remodelling of spheroids led to changes in both cellular and extracellular matrix components (ECMs) over time. Some OEC formed internal clusters, while others organized into a peripheral monolayer, stabilized by ECM and pericyte-like cells, with concomitant increase in surface stiffness. Along spheroid culture, OEC switched from an active to a quiescent state, and their endothelial sprouting potential was significantly abrogated, suggesting that immature spheroids may be more therapeutically relevant. Non-adhesive moulds were subsequently used for triggering rapid, one-step, spheroid formation\/fusion into square-shaped patches, with spheroids uniformly interspaced via a thin cell layer. The high surface area, endothelial sprouting potential, and scalability of the developed spheroid-based patches make them stand out as artificial vascular beds for modular engineering of large tissue constructs.<\/jats:p>","DOI":"10.1088\/1758-5090\/abc790","type":"journal-article","created":{"date-parts":[[2020,11,4]],"date-time":"2020-11-04T17:31:47Z","timestamp":1604511107000},"page":"035008","update-policy":"https:\/\/doi.org\/10.1088\/crossmark-policy","source":"Crossref","is-referenced-by-count":19,"title":["Directed self-assembly of spheroids into modular vascular beds for engineering large tissue constructs"],"prefix":"10.1088","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2664-5889","authenticated-orcid":false,"given":"Daniel T O","family":"Carvalho","sequence":"first","affiliation":[]},{"given":"T\u00e1lia","family":"Feij\u00e3o","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6788-1370","authenticated-orcid":false,"given":"Mariana I","family":"Neves","sequence":"additional","affiliation":[]},{"given":"Ricardo M P","family":"da Silva","sequence":"additional","affiliation":[]},{"given":"Cristina C","family":"Barrias","sequence":"additional","affiliation":[]}],"member":"266","published-online":{"date-parts":[[2021,4,2]]},"reference":[{"key":"bfabc790bib1","doi-asserted-by":"publisher","first-page":"eaaw2459","DOI":"10.1126\/sciadv.aaw2459","type":"journal-article","article-title":"Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels","volume":"5","author":"Skylar-Scott","year":"2019","journal-title":"Sci. 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All rights, including for text and data mining, AI training, and similar technologies, are reserved.","name":"copyright_information","label":"Copyright Information"},{"value":"2019-05-10","name":"date_received","label":"Date Received","group":{"name":"publication_dates","label":"Publication dates"}},{"value":"2020-11-04","name":"date_accepted","label":"Date Accepted","group":{"name":"publication_dates","label":"Publication dates"}},{"value":"2021-04-02","name":"date_epub","label":"Online publication date","group":{"name":"publication_dates","label":"Publication dates"}}]}}