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The current approaches require a strict process of manual inclusion in animal-derived matrix Matrigel\u00ae and are challenged by unpredictability, operators\u2019 skill and expertise, elevated costs, and restricted scalability, impeding their extensive applicability and translational potential. In this study, we present a novel method to generate brain organoids that address these limitations. Our approach does not require a manual, operator-dependent embedding. Instead, it employs a chemically defined hydrogel in which the Matrigel\u00ae is diluted in a solution enriched with sodium alginate (SA) and sodium carboxymethylcellulose (CMC) and used as a bioink to print neural embryoid bodies (nEBs). Immunohistochemical, immunofluorescence, and gene expression analyses confirmed that SA-CMC-Matrigel\u00ae hydrogel can sustain the generation of iPSC-derived cortical cORGs as the conventional Matrigel\u00ae-based approach does. By day 40 of differentiation, hydrogel-based 3D-bioprinted cORGs showed heterogeneous and consistent masses, with a cytoarchitecture resembling an early-stage developmental fetal brain composed of neural progenitor cells PAX6+\/Ki67+ organized into tubular structures, and densely packed cell somas with extensive neurites SYP+, suggestive of cortical tissue-like neuronal layer formation.<\/jats:p>","DOI":"10.3390\/gels11040284","type":"journal-article","created":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T05:38:26Z","timestamp":1744349906000},"page":"284","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Three-Dimensional-Bioprinted Embedded-Based Cerebral Organoids: An Alternative Approach for Mini-Brain In Vitro Modeling Beyond Conventional Generation Methods"],"prefix":"10.3390","volume":"11","author":[{"given":"Rosalba Monica","family":"Ferraro","sequence":"first","affiliation":[{"name":"\u201cAngelo 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