Transcriptome Analysis of Signaling Pathways in Caco-2 Cells Involved in the Formation of Intestinal Villi
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UCERS, BIOLOGY, SELECTION, AND GENE ENGINEERING
Transcriptome Analysis of Signaling Pathways in Caco-2 Cells Involved in the Formation of Intestinal Villi S. V. Nikulina, b, *, M. P. Raigorodskayaa, and D. A. Sakharova aBioclinicum bFar
Scientific Research Center, Moscow, 115088 Russia Eastern Federal University, Vladivostok, 690091 Russia *e-mail: [email protected]
Received October 23, 2019; revised November 10, 2019; accepted January 15, 2020
Abstract—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. Keywords: impedance spectroscopy, intestine, villi, TEER, BMP, PDGF DOI: 10.1134/S0003683820090069
INTRODUCTION In vitro models of the intestinal barrier are widely used in clinical trials to study drug absorption [1]. Barrier-forming epithelial cells are primarily characterized by the ability to generate tight intercellular junctions that separate the apical and basolateral compartments. Monitoring the integrity of these tight junctions is a key task with in vitro models of barrier tissues. Today, there are several methods to control the state of tight junctions; the use of labeled substrates [2, 3] and measurement of the transepithelial/transendothelial resistance (TEER) [4] are the most popular. We showed earlier that impedance spectroscopy for TEER evaluation allows a more reliable identification of defects in obtained models than the classical method. In addition, the suggested method is less laborious and time-consuming than the use of labeled substrates [5]. It is also important to strive for the most complete reproduction of the intestinal structure in the creation of in vitro models. Villi are one of the characteristic features of a fully developed intestine [6]. The mechanism of villus formation is not entirely clear; however, it was shown that the BMP and PDGF signaling pathAbbreviations: BMP—bone morphogenetic protein; FBS—fetal bovine serum; PDGF—platelet derived growth factor; R—Pearson’s correlation coefficient; TEER—transepithelial/transendothelial resistance.
ways play an important role in this process [9, 10]. It was recently show
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