Publication: Establishment of Intestinal Epithelial Cell Monolayers and Their Use in Calcium Switch Assay for Assessment of Intestinal Tight Junction Assembly
Issued Date
2021-01-01
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ISSN
19406029
10643745
10643745
Other identifier(s)
2-s2.0-85109524305
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Mahidol University
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SCOPUS
Bibliographic Citation
Methods in Molecular Biology. Vol.2367, (2021), 273-290
Suggested Citation
Pawin Pongkorpsakol, Wilasinee Satianrapapong, Preedajit Wongkrasant, Peter R. Steinhagen, Nuttha Tuangkijkul, Nutthapoom Pathomthongtaweechai, Chatchai Muanprasat Establishment of Intestinal Epithelial Cell Monolayers and Their Use in Calcium Switch Assay for Assessment of Intestinal Tight Junction Assembly. Methods in Molecular Biology. Vol.2367, (2021), 273-290. doi:10.1007/7651_2021_347 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/76360
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Title
Establishment of Intestinal Epithelial Cell Monolayers and Their Use in Calcium Switch Assay for Assessment of Intestinal Tight Junction Assembly
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.