| Literature DB >> 33861461 |
Pawin Pongkorpsakol1, Wilasinee Satianrapapong2, Preedajit Wongkrasant3, Peter R Steinhagen4, Nuttha Tuangkijkul5, Nutthapoom Pathomthongtaweechai6, Chatchai Muanprasat6.
Abstract
Intestinal barrier function relies primarily on the assembly and integrity of tight junctions, which forms a size-selective barrier. This barrier restricts paracellular movement of solutes in various types of epithelia. Of note, extracellular Ca2+ concentration affects tight junction assembly. Therefore, the removal and re-addition of Ca2+ into cell culture medium of cultured intestinal epithelial cells causes destabilization and reassembly of tight junction to membrane periphery near apical surface, respectively. Based on this principle, the Ca2+-switch assay was established to investigate tight junction assembly in fully differentiated intestinal epithelial cells. This chapter provides a stepwise protocol for culture of intestinal epithelial cell monolayers using T84 cell line as an in vitro model and the Ca2+-switch assay for evaluating tight junction assembly.Entities:
Keywords: Barrier function; Ca2+-switch assay; Epithelial cells; Paracellular permeability; Tight junction; Transepithelial electrical resistance (TER)
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Year: 2021 PMID： 33861461 DOI： 10.1007/7651_2021_347
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745