Caco-2 cells are traditionally used to construct in vitro models of the intestinal barrier. One characteristic of the mature intestine is the presence of villi—connective tissue outgrowths covered with epithelial cells. It was recently shown that Caco-2 cells form structures resembling intestinal villi during prolonged cultivation. In this work, we showed via transcriptome analysis that the BMP and PDGF signaling cascades involved in the formation of villi in vivo are significantly altered during the differentiation of Caco-2 cells and, therefore, can participate in the formation of similar structures in vitro. In particular, we found a significant decrease in the expression of the BMP4, BMP7, and BMP8A genes in differentiated cells as compared to undifferentiated cells. We also first discovered periodic fluctuations in transepithelial resistance upon the differentiation of Caco-2 cells. The period of observed fluctuations indicates that they can occur as a result of cell proliferation during villus formation.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Shah, P., Jogani, V., Bagchi, T., and Misra, A., Role of Caco-2 cell monolayers in prediction of intestinal drug absorption, Biotechnol. Prog., 2006, vol. 22, pp. 186–198.
Hidalgo, I.J., Raub, T.J., and Borchardt, R.T., Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability, Gastroenterology, 1989, vol. 96, pp. 736–749.
Hubatsch, I., Ragnarsson, E.G.E., and Artursson, P., Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers, Nat. Protoc., 2007, vol. 2, pp. 2111–2119.
Srinivasan, B., Kolli, A.R., Esch, M.B., et al., TEER measurement techniques for in vitro barrier model systems, J. Lab. Autom., 2015, vol. 20, pp. 107–126.
Nikulin, S.V., Gerasimenko, T.N., Shilin, S.A., et al., Application of impedance spectroscopy for the control of the integrity of in vitro models of barrier tissues, Bull. Exp. Biol. Med., 2019, vol. 166, pp. 512–516.
Shyer, A.E., Tallinen, T., Nerurkar, N.L., et al., Villification: how the gut gets its villi, Science, 2013, vol. 342, pp. 212–218.
van der Helm, M.W., Henry, O.Y.F., Bein, A., et al., Non-invasive sensing of transepithelial barrier function and tissue differentiation in organs-on-chips using impedance spectroscopy, Lab. Chip, 2019, vol. 19, pp. 452–463.
Nikulin, S.V., Knyazev, E.N., Poloznikov, A.A., et al., Expression of SLC30A10 and SLC23A3 transporter mRNAs in Caco-2 cells correlates with an increase in the area of the apical membrane, Mol. Biol., 2018, vol. 52, pp. 577–582.
Walton, K.D., Freddo, A.M., Wang, S., and Gumucio, D.L., Generation of intestinal surface: an absorbing tale, Development, 2016, vol. 143, pp. 2261–2272.
Chin, A.M., Hill, D.R., Aurora, M., and Spence, J.R., Morphogenesis and maturation of the embryonic and postnatal intestine, Semin. Cell Dev. Biol., 2017, vol. 66, pp. 81–93.
Nikulin, S.V., Knyazev, E.N., Gerasimenko, T.N., et al., Impedance spectroscopy and transcriptome analysis of choriocarcinoma BeWo b30 as a model of human placenta, Mol. Biol., 2019, vol. 53, pp. 411–418.
Nikulin, S.V., Knyazev, E.N., Gerasimenko, T.N., et al., Non-invasive evaluation of extracellular matrix formation in the intestinal epithelium, Bull. Exp. Biol. Med., 2018, vol. 166, pp. 35–38.
Samatov, T.R., Senyavina, N.V., Galatenko, V.V., et al., Tumour-like druggable gene expression pattern of CaCo2 cells in microfluidic chip, BioChip J., 2016, vol. 10, pp. 215–220.
Semenova, O.V, Petrov, V.A., Gerasimenko, T.N., et al., Effect of circulation parameters on functional status of heparg spheroids cultured in microbioreactor, Bull. Exp. Biol. Med., 2016, vol. 161, pp. 425–429.
Henry, O.Y.F., Villenave, R., Cronce, M.J., et al., Organs-on-chips with integrated electrodes for trans-epithelial electrical resistance (TEER) measurements of human epithelial barrier function, Lab. Chip, 2017, vol. 17, pp. 2264–2271.
Izumo, M., Johnson, C.H., and Yamazaki, S., Circadian gene expression in mammalian fibroblasts revealed by real-time luminescence reporting: Temperature compensation and damping, Proc. Natl. Acad. Sci. U. S. A., 2003, vol. 100, pp. 16089–16094.
Izumo, M., Sato, T.R., Straume, M., and Johnson, C.H., Quantitative analyses of circadian gene expression in mammalian cell cultures, PLoS Comput. Biol., 2006, vol. 2, p. e136.
Brown, S.A., Fleury-Olela, F., Nagoshi, E., et al., The period length of fibroblast circadian gene expression varies widely among human individuals, PLoS Biol., 2005, vol. 3, p. e338.
Bairoch, A., The Cellosaurus, a cell-line knowledge resource, J. Biomol. Tech., 2018, vol. 29, pp. 25–38.
The study was funded by the Russian Science Foundation (project no.16-19-10597).
The authors declare that they have no conflicts of interest.
This article does not contain any studies involving animals performed by any of the authors.
This article does not contain any studies involving human participants performed by any of the authors.
Translated by I. Gordon
Abbreviations: BMP—bone morphogenetic protein; FBS—fetal bovine serum; PDGF—platelet derived growth factor; R—Pearson’s correlation coefficient; TEER—transepithelial/transendothelial resistance.
About this article
Cite this article
Nikulin, S.V., Raigorodskaya, M.P. & Sakharov, D.A. Transcriptome Analysis of Signaling Pathways in Caco-2 Cells Involved in the Formation of Intestinal Villi. Appl Biochem Microbiol 56, 898–901 (2020). https://doi.org/10.1134/S0003683820090069
- impedance spectroscopy