scholarly journals Morphofunctional properties of a differentiated Caco2/HT-29 co-culture as an in vitro model of human intestinal epithelium

2018 ◽  
Vol 38 (2) ◽  
Author(s):  
Anita Ferraretto ◽  
Michela Bottani ◽  
Paola De Luca ◽  
Laura Cornaghi ◽  
Francesca Arnaboldi ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
In Vitro Model ◽  
Transepithelial Transport ◽  
Intestinal Cell ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Vitro Model ◽  
Human Intestinal Epithelium ◽  
Ht 29

An intestinal 70/30 Caco2/HT-29 co-culture was set up starting from the parental populations of differentiated cells to mimic the human intestinal epithelium. Co-culture was harvested at confluence 0 (T0) and at 3, 6, 10, and 14 days post confluence after plating (T3, T6, T10, and T14, respectively) for morphological and functional analysis. Transmission electron microscopy revealed different features from T0 to T14: microvilli and a complete junctional apparatus from T6, mucus granules from T3, as also confirmed by PAS/Alcian Blue staining. The specific activity of alkaline phosphatase (ALP), aminopeptidase N (APN), and dipeptidyl peptidase IV (DPPIV) progressively increased after T0, indicating the acquirement of a differentiated and digestive phenotype. Transepithelial electrical resistance (TEER), indicative of the barrier properties of the monolayer, increased from T0 up to T6 reaching values very similar to the human small intestine. The apparent permeability coefficient for Lucifer Yellow (LY), along with morphological analysis, reveals a good status of the tight junctions. At T14, HT-29 cells reduced to 18.4% and formed domes, indicative of transepithelial transport of nutrients. This Caco2/HT-29 co-culture could be considered a versatile and suitable in vitro model of human intestinal epithelium for the presence of more than one prevalent intestinal cell type, by means of a minimum of 6 to a maximum of 14 post-confluence days obtained without the need of particular inducers of subclones and growth support to reach an intestinal differentiated phenotype.

Anti-inflammatory effects of dietary phenolic compounds in an in vitro model of inflamed human intestinal epithelium

2010 ◽  
Vol 188 (3) ◽  
pp. 659-667 ◽  
Author(s):  
Thérèse Sergent ◽  
Neil Piront ◽  
Julie Meurice ◽  
Olivier Toussaint ◽  
Yves-Jacques Schneider
Keyword(s):  
Phenolic Compounds ◽  
Intestinal Epithelium ◽  
In Vitro Model ◽  
Anti Inflammatory ◽  
Vitro Model ◽  
Human Intestinal Epithelium

scholarly journals Food Contaminants Effects on an In Vitro Model of Human Intestinal Epithelium

Toxics ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 135
Author(s):  
Marion Guibourdenche ◽  
Johanna Haug ◽  
Noëllie Chevalier ◽  
Madeleine Spatz ◽  
Nicolas Barbezier ◽  
...  
Keyword(s):  
Gene Expression ◽  
Intestinal Permeability ◽  
Intestinal Epithelium ◽  
In Vitro Model ◽  
Food Contaminants ◽  
Gut Barrier ◽  
Gut Barrier Function ◽  
Vitro Model ◽  
Human Intestinal Epithelium

Pesticide residues represent an important category of food contaminants. Furthermore, during food processing, some advanced glycation end-products resulting from the Maillard reaction can be formed. They may have adverse health effects, in particular on the digestive tract function, alone and combined. We sought to validate an in vitro model of the human intestinal barrier to mimic the effects of these food contaminants on the epithelium. A co-culture of Caco-2/TC7 cells and HT29-MTX was stimulated for 6 h with chlorpyrifos (300 μM), acrylamide (5 mM), Nε-Carboxymethyllysine (300 μM) alone or in cocktail with a mix of pro-inflammatory cytokines. The effects of those contaminants on the integrity of the gut barrier and the inflammatory response were analyzed. Since the co-culture responded to inflammatory stimulation, we investigated whether this model could be used to evaluate the effects of food contaminants on the human intestinal epithelium. CPF alone affected tight junctions’ gene expression, without inducing any inflammation or alteration of intestinal permeability. CML and acrylamide decreased mucins gene expression in the intestinal mucosa, but did not affect paracellular intestinal permeability. CML exposure activated the gene expression of MAPK pathways. The co-culture response was stable over time. This cocktail of food contaminants may thus alter the gut barrier function.

Vitamin D-regulated calcium transport in Caco-2 cells: unique in vitro model

1991 ◽  
Vol 260 (2) ◽  
pp. G207-G212 ◽  
Author(s):  
A. R. Giuliano ◽  
R. J. Wood
Keyword(s):  
Vitamin D ◽  
Cell Line ◽  
Calcium Transport ◽  
In Vitro Model ◽  
Colon Adenocarcinoma ◽  
Intestinal Cell ◽  
Maximal Velocity ◽  
Dependent Manner ◽  
Vitro Model

The human colon adenocarcinoma cell line Caco-2 is the only intestinal cell line to differentiate spontaneously in culture exhibiting structural and biochemical characteristics of mature enterocytes and to possess a vitamin D receptor in the fully differentiated state. Transepithelial calcium transport was characterized in differentiated Caco-2 cells grown on permeable filters supports to assess the potential utility of this cell line as an in vitro model to study 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]-induced calcium transport. Calcium transport was increased in a dose-dependent manner by 1,25(OH)2D3. Total calcium transport at different calcium concentrations could be fitted to a modified Michaelis-Menten equation containing a linear transport component. The maximum rate of saturable calcium transport was increased by 4.3-fold (P less than 0.005) in cells treated with 10(-8) M 1,25(OH)2D3. This treatment also increased the apparent buffer calcium concentration that results in half-maximal velocity from 0.4 to 1.3 mM but had no significant effect on nonsaturable calcium transport. Caco-2 cells grown on permeable filter supports provide a unique in vitro human cell culture model to study the mechanism of vitamin D-regulated transepithelial intestinal calcium transport.

scholarly journals Characterizing Phage-Host Interactions in a Simplified Human Intestinal Barrier Model

2020 ◽  
Vol 8 (9) ◽  
pp. 1374
Author(s):  
María A. Núñez-Sánchez ◽  
Joan Colom ◽  
Lauren Walsh ◽  
Colin Buttimer ◽  
Andrei Sorin Bolocan ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
In Vitro Model ◽  
Cell Model ◽  
Layer Structure ◽  
Intestinal Barrier ◽  
Host Interactions ◽  
Intestinal Epithelia ◽  
Vitro Model ◽  
Barrier Model

An intestinal epithelium model able to produce mucus was developed to provide an environment suitable for testing the therapeutic activity of gut bacteriophages. We show that Enterococcus faecalis adheres more effectively in the presence of mucus, can invade the intestinal epithelia and is able to translocate after damaging tight junctions. Furthermore, Enterococcus phage vB_EfaM_A2 (a member of Herelleviridae that possesses virion associated immunoglobin domains) was found to translocate through the epithelium in the presence and absence of its host bacteria. Phage A2 protected eukaryotic cells by reducing mortality and maintaining the structure of the cell layer structure. We suggest the mammalian cell model utilized within this study as an adaptable in vitro model that can be employed to enable a better understanding of phage–bacteria interactions and the protective impact of phage therapy relating to the intestinal epithelium.

Toxicity, uptake, and nuclear translocation of ingested micro-nanoplastics in an in vitro model of the small intestinal epithelium

2021 ◽  
pp. 112609
Author(s):  
Glen M. DeLoid ◽  
Xiaoqiong Cao ◽  
Dimitrios Bitounis ◽  
Dilpreet Singh ◽  
Paula Montero Llopis ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
In Vitro Model ◽  
Nuclear Translocation ◽  
Small Intestinal Epithelium ◽  
Small Intestinal ◽  
Vitro Model
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