{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T16:01:28Z","timestamp":1790438488716,"version":"4.1.0"},"reference-count":71,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:00:00Z","timestamp":1789516800000},"content-version":"vor","delay-in-days":15,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Pluripotent stem cell (PSC)\u2013derived telencephalic organoids have long been established as an\n                    <jats:italic>in vitro<\/jats:italic>\n                    cell model to study the molecular and cellular features of the developing cerebral cortex and associated brain structures. Numerous directed organoid differentiation protocols have been developed that recapitulate cell subtype specification, cytoarchitectural organization, and cellular function with a high degree of fidelity. Protocol development has focused on the scalability of this\n                    <jats:italic>in vitro<\/jats:italic>\n                    system, and most organoid culture platforms involve a population\u2010based approach in large vessel sizes. This makes comparative study with multiple cell lines both costly and time\u2010consuming, and lowers the throughput for phenotypic screening. We have established a high\u2010throughput approach for\n                    <jats:italic>in vitro<\/jats:italic>\n                    generation of hiPSC\u2010derived telencephalic organoids that can be used to develop comparative phenotypic assays involving high\u2010content imaging platforms. This article first highlights the use of a scaled\u2010down, miniaturized cell culture volume that is amenable to high\u2010throughput production of multiple 384\u2010well plates. Then, using controlled morphogen cues, 3D hiPSC aggregates are directed into an anterior telencephalic identity and further specified into dorsal pallium or ventral subpallium cell fates, which can additionally be combined into assembloids for modeling excitatory and inhibitory networks or left to develop separately as single organoid subtypes. Finally, we introduce a Support Protocol detailing an\n                    <jats:italic>in vitro<\/jats:italic>\n                    3D cell painting assay for organoid morpho\u2010phenotypic characterization. \u00a9 2026 Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : High\u2010throughput 3D aggregation of hiPSCs in morphogen\u2010rich media\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Recombinant inhibitory protein\u2013directed induction of telencephalic organoids\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Morphogen\u2010guided induction of dorsal telencephalic\/pallial organoids\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 4<\/jats:bold>\n                    : Morphogen\u2010guided induction of ventral telencephalic\/subpallial organoids\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol<\/jats:bold>\n                    : 3D live cell painting for organoid morphometric phenotyping\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70449","type":"journal-article","created":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T15:30:29Z","timestamp":1789572629000},"source":"Crossref","is-referenced-by-count":0,"title":["Morphogen\u2010Directed, High\u2010Throughput Development of hiPSC\u2010Derived Telencephalic Organoids for Comparative\n                    <i>In Vitro<\/i>\n                    Phenotyping"],"prefix":"10.1002","volume":"6","author":[{"given":"Dosh","family":"Whye","sequence":"first","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erika M.","family":"Norabuena","sequence":"additional","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wardiya","family":"Afshar\u2010Saber","sequence":"additional","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"},{"name":"Department of Neurology Harvard Medical School  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Becca","family":"Lewis","sequence":"additional","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taryn J.","family":"Polanco","sequence":"additional","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Delaney","family":"Wood","sequence":"additional","affiliation":[{"name":"Department of Neurosciences Stanford University School of Medicine  Stanford California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mustafa","family":"Sahin","sequence":"additional","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"},{"name":"Department of Neurology Harvard Medical School  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth D.","family":"Buttermore","sequence":"additional","affiliation":[{"name":"Human Neuron Core, Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center Boston Children's Hospital Boston Massachusetts"},{"name":"F.M. Kirby Neurobiology Department Boston Children's Hospital  Boston Massachusetts"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,16]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/advs.202515887"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2024.10.016"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2023.08.009"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.117093"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2023.11.013"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596\u2010024\u201001093\u2010w"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4304"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1504393112"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.02.050"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2023.08.013"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.98096"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010020\u20101962\u20100"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature22330"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2021.11.011"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010024\u201002412\u20105"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.105"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms1817"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1529"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010021\u201001126\u20102"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2023.09.003"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41537\u2010022\u201000326\u20109"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2024.1403734"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5128\u201010.2011"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abf5546"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2008.09.002"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1101\/2025.10.01.679721"},{"key":"e_1_2_11_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2020.04.004"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.12.012"},{"key":"e_1_2_11_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41567\u2010022\u201001822\u20106"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abn4705"},{"key":"e_1_2_11_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010025\u201009151\u20103"},{"key":"e_1_2_11_33_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1315710110"},{"key":"e_1_2_11_34_1","doi-asserted-by":"crossref","unstructured":"Kelkar G. 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