{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T04:56:21Z","timestamp":1790571381074,"version":"4.1.0"},"reference-count":52,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100020046","name":"Khalifa Bin Zayed Al Nahyan Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100020046","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Pluripotent stem cell-derived alpha cells (SC-\u03b1 cells) represent a promising component of next-generation cell therapies designed to more fully recapitulate human islet function; however, heterogeneity and the presence of undesired side populations limit their functional maturity and therapeutic potential. Here, we identify the cell surface protein CD98hc (SLC3A2), that when highly expressed in precursor cells, is associated with differentiation toward a more transcriptionally and functionally mature population of human SC-\u03b1 cells.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Directed differentiation was used to generate SC-\u03b1 cells from pluripotent stem cells, which were assessed via single cell RNA sequencing, flow cytometry, and immunofluorescence imaging. Following magnetic activated cell sorting (MACS) of CD98hc enriched precursors, SC-\u03b1 cell function was assessed using glucose stimulated glucagon secretion (GSGS).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We show that CD98hc is upregulated in SC-\u03b1 cells expressing high levels of genes known to play important roles in \u03b1 cell identity and function, and that CD98hc expression is elevated at the pre-\u03b1 precursor stage relative to non\u2013\u03b1 cell populations. We demonstrate that magnetic enrichment of CD98hc\u207a pre-\u03b1 cells derived from multiple pluripotent stem cell lines selectively increases the proportion of the highest CD98hc expressing glucagon-copositive precursors while depleting multiple undesired side populations, including somatostatin-positive and SOX9-positive cells. Although total \u03b1 cell yield equalizes following complete differentiation, SC-\u03b1 cells derived from high-CD98hc enriched precursors exhibit significantly enhanced glucose-responsive glucagon secretion compared to sham-sorted controls, indicating improved functional maturation of this precursor population.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>Together, these findings establish that high expression levels of CD98hc marks a functionally superior \u03b1 cell precursor, that enrichment at the precursor stage can bias differentiation toward higher functional \u03b1 cell output, that stem cell-derived \u03b1 cell populations harbor intrinsic functional heterogeneity, and that selective targeting of precursor populations represents a generalizable strategy to improve the quality and performance of stem cell-derived endocrine cell products.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fendo.2026.1923474","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T04:29:33Z","timestamp":1790569773000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["CD98hc marks a precursor of human stem cell-derived alpha cells primed for functional maturation"],"prefix":"10.3389","volume":"17","author":[{"given":"Sean","family":"Lewis-Brinkman","sequence":"first","affiliation":[{"name":"Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic","place":["Rochester, MN, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiana","family":"Salomon","sequence":"additional","affiliation":[{"name":"Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic","place":["Rochester, MN, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyle R.","family":"Knofczynski","sequence":"additional","affiliation":[{"name":"Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic","place":["Rochester, MN, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caden","family":"Duffy","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biomedical Engineering, Mayo Clinic","place":["Rochester, MN, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quinn P.","family":"Peterson","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biomedical Engineering, Mayo Clinic","place":["Rochester, MN, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/S2213-8587(22)00218-2","article-title":"Global incidence, prevalence, and mortality of type 1 diabetes in 2021 with projection to 2040: a modelling study","volume":"10","author":"Gregory","year":"2022","journal-title":"Lancet Diabetes Endocrinol"},{"key":"B2","unstructured":"International Diabetes Federation"},{"key":"B3","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1001\/jama.2014.16425","article-title":"Estimated life expectancy in a Scottish cohort with type 1 diabetes 2008-2010","volume":"313","author":"Livingstone","year":"2015","journal-title":"JAMA"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.2522\/ptj.20080020","article-title":"Epidemiology of diabetes and diabetes-related complications","volume":"88","author":"Deshpande","year":"2008","journal-title":"Phys Ther"},{"key":"B5","volume-title":"OPTN\/SRTR 2024 Annual Data Report: Deceased Organ Donation","author":"Israni","year":"2014"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1016\/j.transproceed.2022.03.059","article-title":"The 2022 international pancreas transplant registry report-A review","volume":"54","author":"Gruessner","year":"2022","journal-title":"Transplant Proc"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.diabet.2026.101738","article-title":"Overview of the major clinical trials investigating stem cells-based therapies for diabetes","volume":"52","author":"Lin","year":"2026","journal-title":"Diabetes Metab"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.09.040","article-title":"Generation of functional human pancreatic beta cells in vitro","volume":"159","author":"Pagliuca","year":"2014","journal-title":"Cell"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3033","article-title":"Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells","volume":"32","author":"Rezania","year":"2014","journal-title":"Nat Biotechnol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.15252\/embj.201591058","article-title":"Controlled induction of human pancreatic progenitors produces functional beta-like cells in vitro","volume":"34","author":"Russ","year":"2015","journal-title":"EMBO J"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-022-01219-z","article-title":"Functional, metabolic and transcriptional maturation of human pancreatic islets derived from stem cells","volume":"40","author":"Balboa","year":"2022","journal-title":"Nat Biotechnol"},{"key":"B12","doi-asserted-by":"publisher","first-page":"5758","DOI":"10.1038\/s41467-025-60831-0","article-title":"Pancreatic islet beta-cell subtypes are derived from biochemically-distinct and nutritionally-regulated islet progenitors","volume":"16","author":"Brown","year":"2025","journal-title":"Nat Commun"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms11756","article-title":"Human islets contain four distinct subtypes of beta cells","volume":"7","author":"Dorrell","year":"2016","journal-title":"Nat Commun"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/s42255-024-01097-6","article-title":"Exploring pancreatic beta-cell subgroups and their connectivity","volume":"6","author":"Rutter","year":"2024","journal-title":"Nat Metab"},{"key":"B15","doi-asserted-by":"publisher","first-page":"792","DOI":"10.1038\/s41556-019-0316-3","article-title":"Author Correction: Recapitulating endocrine cell clustering in culture promotes maturation of human stem-cell-derived beta cells","volume":"21","author":"Nair","year":"2019","journal-title":"Nat Cell Biol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1168-5","article-title":"Charting cellular identity during human in vitro beta-cell differentiation","volume":"569","author":"Veres","year":"2019","journal-title":"Nature"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2019.11.018","article-title":"Abnormal regulation of glucagon secretion by human islet alpha cells in the absence of beta cells","volume":"50","author":"Liu","year":"2019","journal-title":"EBioMedicine"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.79.23.7322","article-title":"Glucose-induced insulin release depends on functional cooperation between islet cells","volume":"79","author":"Pipeleers","year":"1982","journal-title":"Proc Natl Acad Sci United States America-Biological Sci"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2018.01.015","article-title":"Paracrine interactions within the pancreatic islet determine the glycemic set point","volume":"27","author":"Rodriguez-Diaz","year":"2018","journal-title":"Cell Metab"},{"key":"B20","doi-asserted-by":"publisher","first-page":"2241","DOI":"10.1038\/s41467-020-16049-3","article-title":"A method for the generation of human stem cell-derived alpha cells","volume":"11","author":"Peterson","year":"2020","journal-title":"Nat Commun"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2025.102504","article-title":"Modeling diabetic alpha cell dysfunction using stem cell-derived alpha cells","volume":"20","author":"Shrestha","year":"2025","journal-title":"Stem Cell Rep"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.59912","article-title":"Microtubules regulate pancreatic beta-cell heterogeneity via spatiotemporal control of insulin secretion hot spots","volume":"10","author":"Trogden","year":"2021","journal-title":"Elife"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)36615-3","article-title":"Pancreatic beta cell heterogeneity in glucose-induced insulin secretion","volume":"267","author":"Van Schravendijk","year":"1992","journal-title":"J Biol Chem"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1038\/26246","article-title":"Amino-acid transport by heterodimers of 4F2hc\/CD98 and members of a permease family","volume":"395","author":"Mastroberardino","year":"1998","journal-title":"Nature"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2021.635405","article-title":"Selection for CD26(-) and CD49A(+) cells from pluripotent stem cells-derived islet-like clusters improves therapeutic activity in diabetic mice","volume":"12","author":"Molakandov","year":"2021","journal-title":"Front Endocrinol (Lausanne)"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2022.105056","article-title":"Human alpha cell transcriptomic signatures of types 1 and 2 diabetes highlight disease-specific dysfunction pathways","volume":"25","author":"Bosi","year":"2022","journal-title":"iScience"},{"key":"B27","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1038\/s41598-018-36698-1","article-title":"Considerations for high-yield, high-throughput cell enrichment: fluorescence versus magnetic sorting","volume":"9","author":"Sutermaster","year":"2019","journal-title":"Sci Rep"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2018.05.006","article-title":"Humanised recombinant antibody fragments bind human pancreatic islet cells","volume":"459","author":"Buenafe","year":"2018","journal-title":"J Immunol Methods"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1007\/s002320001017","article-title":"Surface antigen CD98(4F2): not a single membrane protein, but a family of proteins with multiple functions","volume":"173","author":"Deves","year":"2000","journal-title":"J Membr Biol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1016\/j.mam.2012.10.007","article-title":"The SLC3 and SLC7 families of amino acid transporters","volume":"34","author":"Fotiadis","year":"2013","journal-title":"Mol Aspects Med"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-003-1062-7","article-title":"The ancillary proteins of HATs: SLC3 family of amino acid transporters","volume":"447","author":"Palacin","year":"2004","journal-title":"Pflugers Arch"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.2337\/dbi19-0021","article-title":"A primary role for alpha-cells as amino acid sensors","volume":"69","author":"Dean","year":"2020","journal-title":"Diabetes"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2017.05.006","article-title":"Amino acid transporter Slc38a5 controls glucagon receptor inhibition-induced pancreatic alpha cell hyperplasia in mice","volume":"25","author":"Kim","year":"2017","journal-title":"Cell Metab"},{"key":"B34","doi-asserted-by":"publisher","first-page":"74","DOI":"10.2337\/db11-0739","article-title":"Remodeling of hepatic metabolism and hyperaminoacidemia in mice deficient in proglucagon-derived peptides","volume":"61","author":"Watanabe","year":"2012","journal-title":"Diabetes"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-12-281","article-title":"Polyomic profiling reveals significant hepatic metabolic alterations in glucagon-receptor (GCGR) knockout mice: implications on anti-glucagon therapies for diabetes","volume":"12","author":"Yang","year":"2011","journal-title":"BMC Genomics"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.peptides.2023.171039","article-title":"Amino acids and the changing face of the alpha-cell","volume":"166","author":"Hamilton","year":"2023","journal-title":"Peptides"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.2337\/diab.24.8.735","article-title":"Arginine-stimulated acute phase of insulin and glucagon secretion. I. in normal man","volume":"24","author":"Palmer","year":"1975","journal-title":"Diabetes"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1210\/endo-117-3-817","article-title":"Interplay of nutrients and hormones in the regulation of glucagon release","volume":"117","author":"Pipeleers","year":"1985","journal-title":"Endocrinology"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1152\/ajpendo.1990.259.2.E225","article-title":"Role of glucagon in disposal of an amino acid load","volume":"259","author":"Boden","year":"1990","journal-title":"Am J Physiol"},{"key":"B40","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1016\/0168-8278(90)90072-y","article-title":"Glucagon increases hepatic efficacy for urea synthesis","volume":"10","author":"Vilstrup","year":"1990","journal-title":"J Hepatol"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.4161\/isl.3.4.15729","article-title":"Histopaque provides optimal mouse islet purification kinetics: comparison study with Ficoll, iodixanol and dextran","volume":"3","author":"McCall","year":"2011","journal-title":"Islets"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1007\/BF00355463","article-title":"The efficacy of density gradients for islet purification: a comparison of seven density gradients","volume":"3","author":"van Suylichem","year":"1990","journal-title":"Transpl Int"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-009-1259-1","article-title":"A simple two-step protocol for the purification of human pancreatic beta cells","volume":"52","author":"Banerjee","year":"2009","journal-title":"Diabetologia"},{"key":"B44","doi-asserted-by":"publisher","first-page":"299","DOI":"10.2337\/diab.31.4.299","article-title":"Sorting of pancreatic islet cell subpopulations by light scattering using a fluorescence-activated cell sorter","volume":"31","author":"Nielsen","year":"1982","journal-title":"Diabetes"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1210\/endo-117-3-806","article-title":"A new in vitro model for the study of pancreatic A and B cells","volume":"117","author":"Pipeleers","year":"1985","journal-title":"Endocrinology"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.47.12.1974","article-title":"Sorting human beta-cells consequent to targeted expression of green fluorescent protein","volume":"47","author":"Meyer","year":"1998","journal-title":"Diabetes"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1177\/002215540104900412","article-title":"Identification and purification of functional human beta-cells by a new specific zinc-fluorescent probe","volume":"49","author":"Lukowiak","year":"2001","journal-title":"J Histochem Cytochem"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1155\/2008\/165360","article-title":"Pancreatic beta-cell purification by altering FAD and NAD(P)H metabolism","volume":"2008","author":"Smelt","year":"2008","journal-title":"Exp Diabetes Res"},{"key":"B49","doi-asserted-by":"publisher","first-page":"6786","DOI":"10.1038\/s41598-018-24857-3","article-title":"Publisher Correction: Purification of replicating pancreatic beta-cells for gene expression studies","volume":"8","author":"Carballar","year":"2018","journal-title":"Sci Rep"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2026.102828","article-title":"CD133\/CD49a discriminate between human pluripotent stem cell-derived pancreatic beta and alpha cells","volume":"21","author":"Tian","year":"2026","journal-title":"Stem Cell Rep"},{"key":"B51","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1038\/s41587-023-01767-y","article-title":"Dictionary learning for integrative, multimodal and scalable single-cell analysis","volume":"42","author":"Hao","year":"2024","journal-title":"Nat Biotechnol"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1002\/imt2.177","article-title":"ggVennDiagram: Intuitive Venn diagram software extended","volume":"3","author":"Gao","year":"2024","journal-title":"Imeta"}],"container-title":["Frontiers in Endocrinology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2026.1923474\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T04:29:34Z","timestamp":1790569774000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2026.1923474\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,28]]},"references-count":52,"alternative-id":["10.3389\/fendo.2026.1923474"],"URL":"https:\/\/doi.org\/10.3389\/fendo.2026.1923474","relation":{},"ISSN":["1664-2392"],"issn-type":[{"value":"1664-2392","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,28]]},"article-number":"1923474"}}