{"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":1790438488758,"version":"4.1.0"},"reference-count":31,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:00:00Z","timestamp":1790294400000},"content-version":"vor","delay-in-days":24,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"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"}],"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                    The most common treatment for people with people with type 1 diabetes (T1D) requires injections of exogenous insulin to regulate blood glucose homeostasis. A longer\u2010term therapeutic solution for T1D has emerged whereby patients may regain insulin independence through transplantation of donated human pancreatic islets, which contain insulin\u2010secreting beta (\u03b2)\u2010cells. Donated islets, however, are in short supply. Much effort has been dedicated to using human pluripotent stem cell (hPSC)\u2010derived islets (SC\u2010islets) as an unlimited source of insulin\u2010secreting \u03b2\u2010cells. Here, we describe a robust and reproducible 40\u2010day, seven\u2010stage stepwise process for generating hPSC\u2010derived SC\u2010islets. Our differentiation protocol aims to mimic in vivo pancreas development using stage\u2010specific signals to promote the generation of SC\u2010islets. The initial maintenance and the first four stages are performed in two\u2010dimensional monolayer culture; the subsequent three stages require three\u2010dimensional culture in microwells to induce the formation of islet\u2010like clusters. The SC\u2010islets generated at the end of the procedure respond to glucose challenges, reliably express appropriate endocrine markers, contain intracellular insulin, and display an islet\u2010like architecture. Most SC\u2010islets also partially express maturity markers that are often challenging to activate in vitro. These protocols thus provide a scalable platform for generating SC\u2010islets for disease modeling and cell replacement studies. \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Thawing, recovery, and passaging of cryopreserved hPSCs\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : Preparation of solutions and media for hPSC maintenance and expansion\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 2<\/jats:bold>\n                    : Preparation of Matrigel\u2010coated plates for passaging hSPCs\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Differentiation of hPSCs to SC\u2010islets\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70467","type":"journal-article","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T15:55:40Z","timestamp":1790351740000},"source":"Crossref","is-referenced-by-count":0,"title":["A Stepwise Method to Derive Functional Pancreatic Islet\u2010like Clusters from Human Pluripotent Stem Cells"],"prefix":"10.1002","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7029-3385","authenticated-orcid":false,"given":"Manuj","family":"Bandral","sequence":"first","affiliation":[{"name":"Department of Pharmacology University of Michigan Medical School  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jimena","family":"Alvarez\u2010Castanon","sequence":"additional","affiliation":[{"name":"Department of Pharmacology University of Michigan Medical School  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David S.","family":"Lorberbaum","sequence":"additional","affiliation":[{"name":"Department of Pharmacology University of Michigan Medical School  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,25]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.5966\/sctm.2015\u20100079"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587\u2010022\u201001219\u2010z"},{"issue":"2","key":"e_1_2_9_4_1","first-page":"175","article-title":"Transplantation of intact pancreatic islets in rats","volume":"72","author":"Ballinger W. F.","year":"1972","journal-title":"Surgery"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/bs.ctdb.2024.10.007"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2022.101711"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/dom.16501"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010023\u201035716\u20101"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/ddh088"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1259"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596\u2010021\u201000560\u2010y"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.89962"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/244447a0"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2019.02.007"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u2010981\u201015\u20102389\u20102_3"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587\u2010020\u20100492\u20105"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1093\/stcltm\/szab013"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41556\u2010018\u20100271\u20104"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.09.040"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2023.102580"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/ajt.16647"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2506549"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3033"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13287\u2010025\u201004293\u20107"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2013.07.018"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1056\/nejm200007273430401"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa061267"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022\u20103565(25)23964\u20107"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.11.019"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.07.024"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2024.09.004"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010024\u201069645\u20104"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70467","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpz1.70467","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70467","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T15:55:13Z","timestamp":1790438113000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70467"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9]]},"references-count":31,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1002\/cpz1.70467"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70467","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9]]},"article-number":"e70467"}}