000176912 001__ 176912
000176912 005__ 20240229133724.0
000176912 0247_ $$2doi$$a10.3791/62857
000176912 0247_ $$2pmid$$apmid:34570087
000176912 0247_ $$2altmetric$$aaltmetric:114290040
000176912 037__ $$aDKFZ-2021-02154
000176912 041__ $$aEnglish
000176912 082__ $$a570
000176912 1001_ $$aStanifer, Megan L$$b0
000176912 245__ $$aAdapting Gastrointestinal Organoids for Pathogen Infection and Single Cell Sequencing under Biosafety Level 3 (BSL-3) Conditions.
000176912 260__ $$a[S.l.]$$bJoVE$$c2021
000176912 3367_ $$2DRIVER$$aarticle
000176912 3367_ $$2DataCite$$aOutput Types/Journal article
000176912 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1633959120_10606
000176912 3367_ $$2BibTeX$$aARTICLE
000176912 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000176912 3367_ $$00$$2EndNote$$aJournal Article
000176912 500__ $$a#LA:F140#
000176912 520__ $$aHuman intestinal organoids constitute the best cellular model to study pathogen infections of the gastrointestinal tract. These organoids can be derived from all sections of the GI tract (gastric, jejunum, duodenum, ileum, colon, rectum) and, upon differentiation, contain most of the cell types that are naturally found in each individual section. For example, intestinal organoids contain nutrient-absorbing enterocytes, secretory cells (Goblet, Paneth, and enteroendocrine), stem cells, as well as all lineage-specific differentiation intermediates (e.g., early or immature cell types). The greatest advantage in using gastrointestinal tract-derived organoids to study infectious diseases is the possibility of precisely identifying which cell type is targeted by the enteric pathogen and to address whether the different sections of the gastrointestinal tract and their specific cell types similarly respond to pathogen challenges. Over the past years, gastrointestinal models, as well as organoids from other tissues, have been employed to study viral tropism and the mechanisms of pathogenesis. However, utilizing all the advantages of using organoids when employing highly pathogenic viruses represents a technical challenge and requires strict biosafety considerations. Additionally, as organoids are often grown in three dimensions, the basolateral side of the cells is facing the outside of the organoid while their apical side is facing the inside (lumen) of the organoids. This organization poses a challenge for enteric pathogens as many enteric infections initiate from the apical/luminal side of the cells following ingestion. The following manuscript will provide a comprehensive protocol to prepare human intestinal organoids for infection with enteric pathogens by considering the infection side (apical vs. basolateral) to perform single-cell RNA sequencing to characterize cell-type-specific host/pathogen interactions. This method details the preparation of the organoids as well as the considerations needed to perform this work under biosafety level 3 (BSL-3) containment conditions.
000176912 536__ $$0G:(DE-HGF)POF4-316$$a316 - Infektionen, Entzündung und Krebs (POF4-316)$$cPOF4-316$$fPOF IV$$x0
000176912 588__ $$aDataset connected to CrossRef, PubMed, , Journals: inrepo01.inet.dkfz-heidelberg.de
000176912 7001_ $$0P:(DE-He78)4658b59d5b4e54b919fc63ab1213c78f$$aBoulant, Steeve$$b1$$eLast author$$udkfz
000176912 773__ $$0PERI:(DE-600)2259946-0$$a10.3791/62857$$gno. 175, p. 62857$$p62857$$tJoVE$$v175$$x1940-087X$$y2021
000176912 909CO $$ooai:inrepo02.dkfz.de:176912$$pVDB
000176912 9101_ $$0I:(DE-588b)2036810-0$$6P:(DE-He78)4658b59d5b4e54b919fc63ab1213c78f$$aDeutsches Krebsforschungszentrum$$b1$$kDKFZ
000176912 9131_ $$0G:(DE-HGF)POF4-316$$1G:(DE-HGF)POF4-310$$2G:(DE-HGF)POF4-300$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$aDE-HGF$$bGesundheit$$lKrebsforschung$$vInfektionen, Entzündung und Krebs$$x0
000176912 9141_ $$y2021
000176912 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bJOVE-J VIS EXP : 2019$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0320$$2StatID$$aDBCoverage$$bPubMed Central$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bClarivate Analytics Master Journal List$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0160$$2StatID$$aDBCoverage$$bEssential Science Indicators$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)1190$$2StatID$$aDBCoverage$$bBiological Abstracts$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0113$$2StatID$$aWoS$$bScience Citation Index Expanded$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection$$d2021-05-04
000176912 915__ $$0StatID:(DE-HGF)9900$$2StatID$$aIF < 5$$d2021-05-04
000176912 9201_ $$0I:(DE-He78)F140-20160331$$kF140$$lF140 Zelluläre Polarität und virale Infektion$$x0
000176912 980__ $$ajournal
000176912 980__ $$aVDB
000176912 980__ $$aI:(DE-He78)F140-20160331
000176912 980__ $$aUNRESTRICTED