biostudies-literatureMirabelli CEC | Horizon Europe | Excellent Science | HORIZON EUROPE Marie Sklodowska-Curie ActionsHHS | NIH | OSC | Common Fund (NIH Common Fund)UM | Michigan Institute for Clinical and Health ResearchUM | Michigan Institute for Clinical and Health Research (MICHR)NIEHS NIH HHSHHS | NIH | OSC | Common FundEC | Horizon Europe | HORIZON EUROPE Innovative Europee0085522https://www.ebi.ac.uk/biostudies/studies/S-EPMC9683019biostudies-literatureUnknown96(22)Human norovirus (HNoV) accounts for one-fifth of all acute viral gastroenteritis worldwide and an economic burden of ~$60 billion globally. The lack of treatment options against HNoV is in part due to the lack of cultivation systems. Recently, a model of infection in biopsy-derived human intestinal enteroids (HIE) has been described: 3D-HIE are first dispersed in 2D-monolayers and differentiated prior to infection, resulting in a labor-intensive, time-consuming procedure. Here, we present an alternative protocol for HNoV infection of 3D-HIE. We found that 3D-HIE differentiated as efficiently as 2D-monolayers. In addition, immunofluorescence-based quantification of UEA-1, a lectin that stains the villus brush border, revealed that ~80% of differentiated 3D-HIE spontaneously undergo polarityJournal of virologyHuman Norovirus Efficiently Replicates in Differentiated 3D-Human Intestinal Enteroids.PMC9683019812673R01 ES028802P30 ES017885P30ES017885841247UL1TR002240P30DK034933Sexton JZWobus CESantos-Ferreira NRocha-Pereira JGillilland MGNeyts JColacino JACieza RJMirabelli CTaube SfalseHuman Norovirus Efficiently Replicates in Differentiated 3D-Human Intestinal Enteroids.Human norovirus (HNoV) accounts for one-fifth of all acute viral gastroenteritis worldwide and an economic burden of ~$60 billion globally. The lack of treatment options against HNoV is in part due to the lack of cultivation systems. Recently, a model of infection in biopsy-derived human intestinal enteroids (HIE) has been described: 3D-HIE are first dispersed in 2D-monolayers and differentiated prior to infection, resulting in a labor-intensive, time-consuming procedure. Here, we present an alternative protocol for HNoV infection of 3D-HIE. We found that 3D-HIE differentiated as efficiently as 2D-monolayers. In addition, immunofluorescence-based quantification of UEA-1, a lectin that stains the villus brush border, revealed that ~80% of differentiated 3D-HIE spontaneously undergo polarity2022-01-01T00:00:00Z2022 Nov2026-06-14T03:31:38.457Z2025-02-19T03:25:40.852ZS-EPMC96830193634229710.1128/jvi.00855-22