Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties
Abstract Background Traditionally, transformed cell line monolayers have been the standard model for studying epithelial barrier and transport function. Recently, intestinal organoids were proposed as superior in recapitulating the intestine. Typically, 3D organoids are digested and seeded as monola...
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2025-01-01
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| Online Access: | https://doi.org/10.1186/s12915-024-02105-7 |
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| author | Kopano Valerie Masete Dorothee Günzel Jörg-Dieter Schulzke Hans-Jörg Epple Nina A. Hering |
| author_facet | Kopano Valerie Masete Dorothee Günzel Jörg-Dieter Schulzke Hans-Jörg Epple Nina A. Hering |
| author_sort | Kopano Valerie Masete |
| collection | DOAJ |
| description | Abstract Background Traditionally, transformed cell line monolayers have been the standard model for studying epithelial barrier and transport function. Recently, intestinal organoids were proposed as superior in recapitulating the intestine. Typically, 3D organoids are digested and seeded as monolayers on gelatinous matrix pre-coated surfaces for anchorage. As this coat could potentially act as a diffusion barrier, we aimed to generate robust human duodenum-derived organoid monolayers that do not need a gelatinous matrix for anchorage to improve upon existing models to study epithelial transport and barrier function. Results We characterized these monolayers phenotypically regarding polarization, tight junction formation and cellular composition, and functionally regarding uptake of nutrients, ion transport and cytokine-induced barrier dysfunction. The organoid monolayers recapitulated the duodenum phenotypically as well as functionally regarding glucose and short-chain fatty acid uptake. Tumour necrosis factor-alpha induced paracellular transport of 4-kDa Dextran and transcytosis of 44-kDa horseradish peroxidase. Notably, forskolin-stimulated chloride secretion was consistently lower when organoid monolayers were seeded on a layer of basement membrane extract (BME). Conclusions BME-free organoid monolayers represent an improved model for studying transcytotic, paracellular but especially transcellular transport. As BME is extracted from mice, our model furthers efforts to make organoid culture more animal-free. |
| format | Article |
| id | doaj-art-741d325f643b467c92ad232e5719fafb |
| institution | Kabale University |
| issn | 1741-7007 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | BMC Biology |
| spelling | doaj-art-741d325f643b467c92ad232e5719fafb2025-01-12T12:39:52ZengBMCBMC Biology1741-70072025-01-0123111310.1186/s12915-024-02105-7Matrix-free human 2D organoids recapitulate duodenal barrier and transport propertiesKopano Valerie Masete0Dorothee Günzel1Jörg-Dieter Schulzke2Hans-Jörg Epple3Nina A. Hering4Clinical Physiology/Nutritional Medicine, Medical Department, Division of Gastroenterology, Infectiology and Rheumatology, Charité - Universitätsmedizin BerlinClinical Physiology/Nutritional Medicine, Medical Department, Division of Gastroenterology, Infectiology and Rheumatology, Charité - Universitätsmedizin BerlinClinical Physiology/Nutritional Medicine, Medical Department, Division of Gastroenterology, Infectiology and Rheumatology, Charité - Universitätsmedizin BerlinDepartment of Gastroenterology, Rheumatology and Infectious Diseases, Charité - Universitätsmedizin BerlinDepartment of General and Visceral Surgery, Campus Benjamin Franklin, Charité - Universitätsmedizin BerlinAbstract Background Traditionally, transformed cell line monolayers have been the standard model for studying epithelial barrier and transport function. Recently, intestinal organoids were proposed as superior in recapitulating the intestine. Typically, 3D organoids are digested and seeded as monolayers on gelatinous matrix pre-coated surfaces for anchorage. As this coat could potentially act as a diffusion barrier, we aimed to generate robust human duodenum-derived organoid monolayers that do not need a gelatinous matrix for anchorage to improve upon existing models to study epithelial transport and barrier function. Results We characterized these monolayers phenotypically regarding polarization, tight junction formation and cellular composition, and functionally regarding uptake of nutrients, ion transport and cytokine-induced barrier dysfunction. The organoid monolayers recapitulated the duodenum phenotypically as well as functionally regarding glucose and short-chain fatty acid uptake. Tumour necrosis factor-alpha induced paracellular transport of 4-kDa Dextran and transcytosis of 44-kDa horseradish peroxidase. Notably, forskolin-stimulated chloride secretion was consistently lower when organoid monolayers were seeded on a layer of basement membrane extract (BME). Conclusions BME-free organoid monolayers represent an improved model for studying transcytotic, paracellular but especially transcellular transport. As BME is extracted from mice, our model furthers efforts to make organoid culture more animal-free.https://doi.org/10.1186/s12915-024-02105-7Matrigel-freeBarrier functionOrganoidsSmall intestine3Rs Principle |
| spellingShingle | Kopano Valerie Masete Dorothee Günzel Jörg-Dieter Schulzke Hans-Jörg Epple Nina A. Hering Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties BMC Biology Matrigel-free Barrier function Organoids Small intestine 3Rs Principle |
| title | Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties |
| title_full | Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties |
| title_fullStr | Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties |
| title_full_unstemmed | Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties |
| title_short | Matrix-free human 2D organoids recapitulate duodenal barrier and transport properties |
| title_sort | matrix free human 2d organoids recapitulate duodenal barrier and transport properties |
| topic | Matrigel-free Barrier function Organoids Small intestine 3Rs Principle |
| url | https://doi.org/10.1186/s12915-024-02105-7 |
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