TY - JOUR
T1 - A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine
AU - Roodsant, Thomas
AU - Navis, Marit
AU - Aknouch, Ikrame
AU - Renes, Ingrid B.
AU - van Elburg, Ruurd M.
AU - Pajkrt, Dasja
AU - Wolthers, Katja C.
AU - Schultsz, Constance
AU - van der Ark, Kees C. H.
AU - Sridhar, Adithya
AU - Muncan, Vanesa
PY - 2020/6/9
Y1 - 2020/6/9
N2 - Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and Listeria monocytogenes were evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research.
AB - Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and Listeria monocytogenes were evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research.
KW - enteric pathogens
KW - gut barrier
KW - host-pathogen interactions
KW - human organoids
KW - polarized epithelium
KW - small intestine
UR - http://www.scopus.com/inward/record.url?scp=85087024410&partnerID=8YFLogxK
U2 - https://doi.org/10.3389/fcimb.2020.00272
DO - https://doi.org/10.3389/fcimb.2020.00272
M3 - Article
C2 - 32656095
SN - 2235-2988
VL - 10
JO - Frontiers in cellular and infection microbiology
JF - Frontiers in cellular and infection microbiology
M1 - 272
ER -