biostudies-literatureRamani ABundesministerium für Bildung und Forschung (Federal Ministry of Education and Research)Deutsche Forschungsgemeinschaft (German Research Foundation)Volkswagen Foundation (VolkswagenStiftung)Fritz Thyssen Stiftung (Fritz Thyssen Foundation)Deutsche Krebshilfe (German Cancer Aid)10703https://www.ebi.ac.uk/biostudies/studies/S-EPMC11659410biostudies-literatureUnknown15(1)Brain organoids offer unprecedented insights into brain development and disease modeling and hold promise for drug screening. Significant hindrances, however, are morphological and cellular heterogeneity, inter-organoid size differences, cellular stress, and poor reproducibility. Here, we describe a method that reproducibly generates thousands of organoids across multiple hiPSC lines. These High Quantity brain organoids (Hi-Q brain organoids) exhibit reproducible cytoarchitecture, cell diversity, and functionality, are free from ectopically active cellular stress pathways, and allow cryopreservation and re-culturing. Patient-derived Hi-Q brain organoids recapitulate distinct forms of developmental defects: primary microcephaly due to a mutation in CDK5RAP2 and progeria-associated defects oNature communicationsReliability of high-quantity human brain organoids for modeling microcephaly, glioma invasion and drug screening.PMC11659410RU 2795 ("Synapses under Stress"), Ro2327/13-27011427610.20.2.031MNVIP+ (03VP10540)BU 2974/3-2 - SPP2127, EXC-2151-390873048-Cluster of Excellence- ImmunoSensation2Project-ID 503306912 - FOR5547SPP2127 GO 2301/5-2Freigeist-A110720Mariappan ALin SGabriel EMuller SD'Alessandris QGFreifeld LRathinam DRamani AGoureau OJadhav VGopalakrishnan JMirsaidi AHadian KJurisch-Yaksi NAltinisik NBusskamp VGerkau NJRose CRPasquini GVinchure OSMuotri ARicci-Vitiani LWollnik BWindoffer HPallini RRothenaigner IfalseReliability of high-quantity human brain organoids for modeling microcephaly, glioma invasion and drug screening.Brain organoids offer unprecedented insights into brain development and disease modeling and hold promise for drug screening. Significant hindrances, however, are morphological and cellular heterogeneity, inter-organoid size differences, cellular stress, and poor reproducibility. Here, we describe a method that reproducibly generates thousands of organoids across multiple hiPSC lines. These High Quantity brain organoids (Hi-Q brain organoids) exhibit reproducible cytoarchitecture, cell diversity, and functionality, are free from ectopically active cellular stress pathways, and allow cryopreservation and re-culturing. Patient-derived Hi-Q brain organoids recapitulate distinct forms of developmental defects: primary microcephaly due to a mutation in CDK5RAP2 and progeria-associated defects o2024-01-01T00:00:00Z2024 Dec2025-04-18T13:01:30.926Z2025-04-06T22:27:55.533ZS-EPMC116594103970247710.1038/s41467-024-55226-6