Maturation of human intestinal organoids in vitro facilitates colonization by commensal lactobacilli by reinforcing the mucus layer
Abstract
Lactobacilli, which are probiotic commensal bacteria that mainly reside in the human small intestine, have attracted attention for their ability to exert health‐promoting effects and beneficially modulate host immunity. However, host epithelial‐commensal bacterial interactions are still largely unexplored because of limited access to human small intestinal tissues. Recently, we described an in vitro maturation technique for generating adult‐like, mature human intestinal organoids (hIOs) from human pluripotent stem cells (hPSCs) that closely resemble the in vivo tissue structure and cellular diversity. Here, we established an in vitro human model to study the response to colonization by commensal bacteria using luminal microinjection into mature hIOs, allowing for the direct examination of epithelial‐bacterial interactions. Lactobacillus reuteri and Lactobacillus plantarum were more likely to survive and colonize when microinjected into the lumen of mature hIOs than when injected into immature hIOs, as determined by scanning electron microscopy, colony formation assay, immunofluorescence, and real‐time imaging with L plantarum expressing red fluorescent protein. The improved mature hIO‐based host epithelium system resulted from enhanced intestinal epithelial integrity via upregulation of mucus secretion and tight junction proteins. Our study indicates that mature hIOs are a physiologically relevant in vitro model system for studying commensal microorganisms.
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Functional intestinal organoids have a relatively thick mucus layer because mature goblet cells secrete mucus.
“Furtherly, functional intestinal organoids exhibit a relatively thick mucus layer due to mucus secretion from mature goblet cells122, which can be detected by Alcian blue (AB) and periodic acid-Schiff (PAS) staining123,124.”
From Organoids (2022) · cites this paper as [123]
Cites
The gut-brain axis: interactions between enteric microbiota, central and enteric nervous systems.
(2015) · PubMed Central · Cited by 2,540 · see citing works
Gut biogeography of the bacterial microbiota
Donaldson, G. P., Lee, S. M., & Mazmanian, S. K. (2016) · Nature Reviews Microbiology · doi:10.1038/nrmicro3552 · Cited by 2,535 · see citing works
Milani, C., Duranti, S., Bottacini, F., Casey, E., Turroni, F., Mahony, J., Belzer, C., Delgado Palacio, S., Arboleya Montes, S., Mancabelli, L., et al. (2017) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.00036-17 · Cited by 1,950 · see citing works
Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro
Spence, J., Mayhew, C., Rankin, S., Kuhar, M., Vallance, J. E., Tolle, K., Hoskins, E. E., Kalinichenko, V., Wells, S., Zorn, A., et al. (2010) · Nature · doi:10.1038/nature09691 · Cited by 1,850 · see citing works
Intestinal barrier function: molecular regulation and disease pathogenesis
Groschwitz, K. R., & Hogan, S. P. (2009) · Journal of Allergy and Clinical Immunology · doi:10.1016/j.jaci.2009.05.038 · Cited by 1,775 · see citing works
Symbiotic bacteria direct expression of an intestinal bactericidal lectin
Cash, H. L., Whitham, C. V., Behrendt, C. L., & Hooper, L. V. (2006) · Science · doi:10.1126/science.1127119 · Cited by 1,432 · see citing works
Mucins in the mucosal barrier to infection
Lindén, S. K., SUTTON, P. R. N., Karlsson, N. G., Korolik, V., & McGuckin, M. A. (2008) · Mucosal Immunology · doi:10.1038/mi.2008.5 · Cited by 1,241 · see citing works
Antagonistic activities of lactobacilli and bifidobacteria against microbial pathogens
Servin, A. L. (2004) · FEMS Microbiology Reviews · doi:10.1016/j.femsre.2004.01.003 · Cited by 1,234 · see citing works
The microbiome in early life: Implications for health outcomes
Tamburini, S., Shen, N., Wu, H. C., & Clemente, J. C. (2016) · Nature Medicine · doi:10.1038/nm.4142 · Cited by 1,213 · see citing works
The Human Gut Microbiome – A Potential Controller of Wellness and Disease
Kho, Z. Y., & Lal, S. K. (2018) · Frontiers in Microbiology · doi:10.3389/fmicb.2018.01835 · Cited by 1,116 · see citing works
Modelling human development and disease in pluripotent stem-cell-derived gastric organoids
McCracken, K. W., Catá, E. M., Crawford, C. M., Sinagoga, K. L., Schumacher, M., Rockich, B. E., Tsai, Y.-H., Mayhew, C. N., Spence, J. R., Zavros, Y., et al. (2014) · Nature · doi:10.1038/nature13863 · Cited by 980 · cited 1× · see citing works
“17 Though these organoids offer great advantages in modeling host-microbe interactions, their success likely depends on the similarity to their in vivo counterparts.”
BackgroundMucin glycan foraging in the human gut microbiome
Tailford, L. E., Crost, E. H., Kavanaugh, D. W., & Juge, N. (2015) · Frontiers in Genetics · doi:10.3389/fgene.2015.00081 · Cited by 851 · see citing works
Disease modeling in stem cell-derived 3D organoid systems
Dutta, D., Heo, I., & Clevers, H. (2017) · Trends in Molecular Medicine · doi:10.1016/j.molmed.2017.02.007 · Cited by 838 · see citing works
Effects of short chain fatty acids on gut morphology and function
Scheppach, W. (1994) · Gut · doi:10.1136/gut.35.1_suppl.s35 · Cited by 756 · see citing works
Peng, L., He, Z., Chen, W., Holzman, I. R., & Lin, J. (2007) · Pediatric Research · doi:10.1203/01.pdr.0000250014.92242.f3 · Cited by 518 · see citing works
Assembly of the human intestinal microbiota
Dethlefsen, L., Eckburg, P. B., Bik, E. M., & Relman, D. A. (2006) · Trends in Ecology & Evolution · doi:10.1016/j.tree.2006.06.013 · Cited by 509 · see citing works
Hooper, L. V., Xu, J., Falk, P. G., Midtvedt, T., & Gordon, J. I. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.17.9833 · Cited by 500 · see citing works
Pathogenic relevance of Lactobacillus: A retrospective review of over 200 cases
Cannon, J. P., Lee, T. A., Bolanos, J. T., & Danziger, L. H. (2005) · European Journal of Clinical Microbiology & Infectious Diseases · doi:10.1007/s10096-004-1253-y · Cited by 478 · see citing works
Fight them or feed them: How the intestinal mucus layer manages the gut microbiota
Schroeder, B. O. (2019) · Gastroenterology report · doi:10.1093/gastro/goy052 · Cited by 464 · see citing works
Intestinal Lactobacillus in health and disease, a driver or just along for the ride?
Heeney, D. D., Gareau, M. G., & Marco, M. L. (2017) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2017.08.004 · Cited by 367 · see citing works
Leslie, J. L., Huang, S., Opp, J. S., Nagy, M. S., Kobayashi, M., Young, V. B., & Spence, J. R. (2014) · Infection and Immunity · doi:10.1128/iai.02561-14 · Cited by 330 · see citing works
Sengupta, R., Altermann, E., Anderson, R. C., McNabb, W. C., Moughan, P. J., & Roy, N. C. (2013) · Mediators of Inflammation · doi:10.1155/2013/237921 · Cited by 287 · see citing works
Forbester, J. L., Goulding, D., Vallier, L., Hannan, N. R. F., Hale, C., Pickard, D., Mukhopadhyay, S., & Dougan, G. (2015) · Infection and Immunity · doi:10.1128/iai.00161-15 · Cited by 268 · see citing works
Yu, L. C. (2012) · World Journal of Gastrointestinal Pathophysiology · doi:10.4291/wjgp.v3.i1.27 · Cited by 268 · see citing works
Lactobacilli and Bifidobacteria in Human Breast Milk
Soto, A., Martín, V., Jiménez, E., Mader, I., Rodríguez, J. M., & Fernández, L. (2014) · Journal of Pediatric Gastroenterology and Nutrition · doi:10.1097/mpg.0000000000000347 · Cited by 252 · see citing works
Finkbeiner, S. R., Hill, D. R., Altheim, C. H., Dedhia, P. H., Taylor, M. J., Tsai, Y.-H., Chin, A. M., Mahé, M. M., Watson, C. L., Freeman, J. J., et al. (2015) · Stem Cell Reports · doi:10.1016/j.stemcr.2015.04.010 · Cited by 250 · see citing works
The use of Lactobacillus as an alternative of antibiotic growth promoters in pigs: A review
Dowarah, R., Verma, A. K., & Agarwal, N. (2016) · Animal nutrition · doi:10.1016/j.aninu.2016.11.002 · Cited by 225 · see citing works
Castillo, N. A., Perdigón, G., & de LeBlanc, A. D. M. (2011) · BMC Microbiology · doi:10.1186/1471-2180-11-177 · Cited by 211 · see citing works
Zotta, T., Parente, E., & Ricciardi, A. (2017) · Journal of Applied Microbiology · doi:10.1111/jam.13399 · Cited by 202 · see citing works
Hill, D. R., Huang, S., Nagy, M. S., Yadagiri, V. K., Fields, C., Thornhill, D., Bons, B., Dedhia, P. H., Chin, A. M., Tsai, Y.-H., et al. (2017) · eLife · doi:10.7554/elife.29132 · Cited by 173 · see citing works
Intestinal in vitro and ex vivo Models to Study Host-Microbiome Interactions and Acute Stressors
Pearce, S., Coia, H. G., Karl, J. P., Pantoja-Feliciano, I. G., Zachos, N. C., & Racicot, K. (2018) · Frontiers in Physiology · doi:10.3389/fphys.2018.01584 · Cited by 159 · see citing works
Karve, S. S., Pradhan, S. K., Ward, D. V., & Weiss, A. A. (2017) · PLoS ONE · doi:10.1371/journal.pone.0178966 · Cited by 157 · see citing works
A small intestinal organoid model of non-invasive enteric pathogen–epithelial cell interactions
Wilson, S. S., Tocchi, A., Holly, M. K., Parks, W. C., & Smith, J. G. (2014) · Mucosal Immunology · doi:10.1038/mi.2014.72 · Cited by 156 · see citing works
Host–microbe interactions shaping the gastrointestinal environment
Kaiko, G. E., & Stappenbeck, T. S. (2014) · Trends in Immunology · doi:10.1016/j.it.2014.08.002 · Cited by 151 · see citing works
Sheng, Y., Lourie, R., Lindén, S. K., Jeffery, P. L., Roche, D., Tran, T. V., Png, C. W., Waterhouse, N. J., SUTTON, P. R. N., Florin, T. H., et al. (2011) · Gut · doi:10.1136/gut.2011.239194 · Cited by 140 · see citing works
Jung, K. B., Lee, H., Son, Y. S., Lee, M.-O., Kim, Y.-D., Oh, S. J., Kwon, O., Cho, S., Cho, H.-S., Kim, D.-S., et al. (2018) · Nature Communications · doi:10.1038/s41467-018-05450-8 · Cited by 125 · see citing works
Self-citationHuman mini-guts: New insights into intestinal physiology and host–pathogen interactions
In, J. G., Foulke-Abel, J., Estes, M. K., Zachos, N. C., Kovbasnjuk, O., & Donowitz, M. (2016) · Nature Reviews Gastroenterology & Hepatology · doi:10.1038/nrgastro.2016.142 · Cited by 125 · see citing works
Autoimmune host–microbiota interactions at barrier sites and beyond
Ruff, W. E., & Kriegel, M. A. (2015) · Trends in Molecular Medicine · doi:10.1016/j.molmed.2015.02.006 · Cited by 122 · see citing works
Biologically inspired approaches to enhance human organoid complexity
Holloway, E. M., Capeling, M. M., & Spence, J. R. (2019) · Development · doi:10.1242/dev.166173 · Cited by 96 · see citing works
Simpson, M. R., Avershina, E., Storrø, O., Johnsen, R., Rudi, K., & Øien, T. (2017) · Journal of Dairy Science · doi:10.3168/jds.2017-13411 · Cited by 92 · see citing works
Engineered Human Gastrointestinal Cultures to Study the Microbiome and Infectious Diseases
Blutt, S. E., Crawford, S. E., Ramani, S., Zou, W. Y., & Estes, M. K. (2017) · Cellular and Molecular Gastroenterology and Hepatology · doi:10.1016/j.jcmgh.2017.12.001 · Cited by 86 · see citing works
Proteolytic degradation of human salivary MUC5B by dental biofilms
Wickström, C., Herzberg, M. C., Beighton, D., & Svensäter, G. (2009) · Microbiology · doi:10.1099/mic.0.030536-0 · Cited by 85 · see citing works
Tafazoli, F., Magnusson, K., & Zheng, L. (2003) · Infection and Immunity · doi:10.1128/iai.71.2.872-881.2003 · Cited by 83 · see citing works
Lee, M.-O., Jung, K. B., Jo, S.-J., Hyun, S.-A., Moon, K.-S., Seo, J.-W., Kim, S.-H., & Son, M.-Y. (2019) · Journal of Biological Engineering · doi:10.1186/s13036-019-0139-6 · Cited by 72 · see citing works
Self-citationhPSC-derived lung and intestinal organoids as models of human fetal tissue
Aurora, M., & Spence, J. (2016) · Developmental Biology · doi:10.1016/j.ydbio.2016.06.006 · Cited by 65 · see citing works
Intestinal MUC2 Mucin Supramolecular Topology by Packing and Release Resting on D3 Domain Assembly
Nilsson, H. E., Ambort, D., Bäckström, M., et al. (2014) · Journal of Molecular Biology · doi:10.1016/j.jmb.2014.04.027 · Cited by 59 · see citing works
Fuochi, V., Coniglio, M. A., Laghi, L., Rescifina, A., Caruso, M., Stivala, A., & Furneri, P. M. (2019) · Frontiers in Microbiology · doi:10.3389/fmicb.2019.01403 · Cited by 49 · see citing works
Nononcogenic restoration of the intestinal barrier by E. coli–delivered human EGF
Yu, M. R., Kim, J., Ahn, J. H., & Moon, Y. (2019) · JCI Insight · doi:10.1172/jci.insight.125166 · Cited by 48 · see citing works
Self-citationLee, H., Son, Y. S., Lee, M., Ryu, J., Park, K., Kwon, O., Jung, K. B., Kim, K., Ryu, T. Y., Baek, A., et al. (2020) · Theranostics · doi:10.7150/thno.41534 · Cited by 48 · see citing works
Self-citationGut microbiome investigation in celiac disease: from methods to its pathogenetic role
Sacchetti, L., & Nardelli, C. (2019) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2019-0657 · Cited by 46 · see citing works
Recent progress in organoid culture to model intestinal epithelial barrier functions
Nakamura, T. (2018) · International Immunology · doi:10.1093/intimm/dxy065 · Cited by 38 · see citing works
Ret receptor tyrosine kinase sustains proliferation and tissue maturation in intestinal epithelia
Perea, D., Guiu, J., Hudry, B., Konstantinidou, C., Milona, A., Hadjieconomou, D., Carroll, T., Hoyer, N., Natarajan, D., Kallijärvi, J., et al. (2017) · The EMBO Journal · doi:10.15252/embj.201696247 · Cited by 38 · see citing works
Jung, K. B., Lee, H., Son, Y. S., Lee, J. H., Cho, H., Lee, M., Oh, J., Lee, J., Kim, S., Jung, C., et al. (2017) · The FASEB Journal · doi:10.1096/fj.201700504r · Cited by 33 · see citing works
Self-citationJung, K. B., Kwon, O., Lee, M., Lee, H., Son, Y. S., Habib, O., Oh, J., Cho, H., Jung, C., Kim, J., et al. (2019) · Journal of Clinical Medicine · doi:10.3390/jcm8070976 · Cited by 22 · see citing works
Self-citationSon, Y. S., Choi, K., Lee, H., Kwon, O., Jung, K. B., Cho, S., Baek, J., Son, B., Kang, S., Kang, M.-G., et al. (2019) · Stem Cells and Development · doi:10.1089/scd.2018.0181 · Cited by 16 · see citing works
Self-citation3D Human Adipose-Derived Stem Cell Clusters as a Model for In Vitro Fibrosis
Rajangam, T., Park, M. H., & Kim, S. (2016) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2016.0037 · Cited by 14 · see citing works
Self-citationNew Age Strategies To Reconstruct Mucosal Tissue Colonization and Growth in Cell Culture Systems
Fasciano, A. C., Mecsas, J. C., & Isberg, R. R. (2019) · Microbiology Spectrum · doi:10.1128/microbiolspec.bai-0013-2019 · Cited by 13 · see citing works
Lactobacillus porci sp. nov., isolated from small intestine of a swine
Kim, J., Choe, H., Kim, K. M., Lee, Y.-R., Rhee, M.-S., & Park, D. (2018) · INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY · doi:10.1099/ijsem.0.002949 · Cited by 10 · see citing works
Self-citationPromoting intestinal adaptation
Wallis, K., Lloyd, D. A., & Gabe, S. M. (2007) · British Journal of Hospital Medicine · doi:10.12968/hmed.2007.68.1.22648 · Cited by 5 · see citing works
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- Authors
- Son, Y. S., Ki, S. J., Thanavel, R., Kim, J., Lee, M., Kim, J., Jung, C., Han, T., Cho, H., Ryu, C., Kim, S., Park, D., & Son, M.
- Affiliations
- Korea Research Institute of Bioscience and Biotechnology (KR)
- Center for Biomaterials Korea Institute of Science and Technology (KIST) Seoul Republic of Korea
- Korea Institute of Science and Technology (KR)
- Corresponding author
- Kim, Sang‐Heon
- Park, Doo‐Sang
- Son, Mi‐Young
- ORCID
- Venue
- The FASEB Journal
- Publisher
- Wiley
- Published
- 2020-06-30
- Volume
- 34
- Issue
- 8
- Pages
- 9899-9910
- ISSN
- 0892-6638 (ISSN-L)
- 1530-6860
- OpenAlex
- W3038382700
- Cited by
- 45
- References
- 59
- Citations by year
- 2026: 6 · 2025: 3 · 2024: 9 · 2023: 7 · 2022: 11 · 2021: 6 · 2020: 3
- FWCI
- 4.37
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The FASEB Journal (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Research Foundation of Korea (NRF‐2018M3A9H3023077)
- Rural Development Administration (PJ015049)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Digestive system and related health
- Keywords
- human intestinal organoids, organoid maturation, Lactobacillus reuteri, Lactobacillus plantarum, bacterial colonization, mucus layer, tight junction proteins, intestinal epithelial integrity, human pluripotent stem cells, probiotics
- MeSH terms
- Cell Differentiation, Cells, Cultured, Humans, Intestinal Mucosa, Intestines, Lactobacillus, Organoids, Pluripotent Stem Cells, In Vitro Techniques
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