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T84 monolayers are superior to Caco-2 as a model system of colonocytes

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Abstract

Colonic adenocarcinoma-derived Caco-2 and T84 epithelial cell lines are frequently used as in vitro model systems of functional epithelial barriers. Both are utilised interchangeably despite evidence that differentiated Caco-2 cells are more reminiscent of small intestinal enterocytes than of colonocytes, whereas differentiated T84 cells are less well characterised. The aim of this study was, therefore, to further characterise and compare differentiated Caco-2 and T84 cells. The objectives were to (1) compare the brush border morphology, (2) measure the expression of enterocyte- and colonocyte-specific genes and (3) compare their response to butyrate, which is dependent on the monocarboxylate transporter 1 (MCT1), an apical protein expressed primarily in colonocytes. T84 microvilli were significantly shorter than those of Caco-2 cells, which is a characteristic difference between small intestinal enterocytes and colonocytes. Also, enterocyte-associated brush border enzymes expressed in differentiated Caco-2 cells were not increased during T84 maturation, whereas colonic markers such as MCT1 were more abundant in differentiated T84 cells compared to differentiated Caco-2 cells. Consequently, T84 cells displayed a dose-responsive improvement of barrier function towards butyrate, which was absent in Caco-2 cells. On the other hand, differences in epithelial toll-like receptor expression between Caco-2 and T84 monolayers did not result in a corresponding differential functional response. We conclude that differentiated Caco-2 and T84 cells have distinct morphological, biochemical and functional characteristics, suggesting that T84 cells do not acquire the biochemical signature of mature small intestinal enterocytes like Caco-2 cells, but retain much of their original colonic characteristics throughout differentiation. These findings can help investigators select the appropriate intestinal epithelial cell line for specific in vitro research purposes.

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Abbreviations

ALPI:

Alkaline phosphatase, intestinal

ANPEP:

Alanyl aminopeptidase, membrane

CFTR:

Cystic fibrosis transmembrane conductance regulator

DDP4:

Dipeptidyl peptidase 4

DMEM:

Dulbecco’s Modified Eagle Medium

FBS:

Fetal bovine serum

FLG:

Flagellin

HMBS:

Hydroxymethyl-bilane synthase

IL-8:

Interleukin-8

LCT:

Lactase

LPS:

Lipopolysaccharide

MCT1:

Monocarboxylate transporter 1

MGAM:

Maltase-glucoamylase

MS4A12:

Membrane spanning 4-domains A12

NaB:

Sodium butyrate

NRQs:

Normalised relative quantities

OPLS:

Orthogonal partial least squares

PCA:

Principle component analysis

qRT-PCR:

Quantitative real-time PCR

SEM:

Standard error of the mean

SI:

Sucrase-isomaltase

TEER:

Transepithelial electrical resistance

TEM:

Transmission electron microscopy

TLR:

Toll-like receptor

VIP:

Variable importance for the projection.

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Acknowledgements

The authors would like to thank Petra Van Wassenhove, Griet Driesschaert and Jelle De Medts for technical support, and Ran Rumes for electron microscopy.

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Authors and Affiliations

  1. Department of Gastroenterology, Ghent University, De Pintelaan 185, 0MRB2, 9000, Ghent, Belgium

    Sarah Devriese, Lien Van den Bossche, Sophie Van Welden, Tom Holvoet, Pieter Hindryckx, Martine De Vos & Debby Laukens

  2. ProDigest BVBA, Technologiepark 3, 9052, Ghent, Belgium

    Iris Pinheiro

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  1. Sarah Devriese
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  2. Lien Van den Bossche
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  3. Sophie Van Welden
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  4. Tom Holvoet
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  6. Pieter Hindryckx
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  7. Martine De Vos
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  8. Debby Laukens
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Corresponding author

Correspondence to Debby Laukens.

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Funding

This study was funded by the Flemish Agency for Innovation by Science and Technology (Grant Number SBO-100016). LVDB, TH and DL are paid by grants from the Research Fund—Flanders (FWO 11J9915N, 13/ASP/145 and 1298213N).

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The authors declare that they have no conflict of interest.

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Devriese, S., Van den Bossche, L., Van Welden, S. et al. T84 monolayers are superior to Caco-2 as a model system of colonocytes. Histochem Cell Biol 148, 85–93 (2017). https://doi.org/10.1007/s00418-017-1539-7

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