Generation of mouse small intestinal epithelial cell lines that allow the analysis of specific innate immune functions

• Published in: PloS one 2013-08, Vol.8 (8), p.e72700-e72700
• Main Authors: Schwerk, Johannes, Köster, Mario, Hauser, Hansjörg, Rohde, Manfred, Fulde, Marcus, Hornef, Mathias W, May, Tobias
• Format: Article
• Language: English
• Subjects: Animals; Antigens; Bacteria - immunology; Bacteria - pathogenicity; Biological response modifiers; Biomarkers - metabolism; Biotechnology; Cell culture; Cell Line; Cell lines; Cell Polarity - physiology; Cell Proliferation; Cytokines; Drug development; Embryo, Mammalian; Embryos; Epidemiology; Epithelial cells; Epithelium; Genes; Genetic engineering; Homeostasis; Immortalization; Immunity, Innate - physiology; Immunology; Infections; Inflammatory bowel disease; Interferon; Interferon Type I - metabolism; Intestinal Mucosa - cytology; Intestinal Mucosa - embryology; Intestinal Mucosa - immunology; Intestine, Small - cytology; Intestine, Small - embryology; Intestine, Small - immunology; Mice; Microorganisms; Microscopy; Mucosa; Primary Cell Culture - methods; Reproducibility; Rodents; Small intestine; Transgenic animals; Transgenic mice; Viruses; Viruses - immunology; Viruses - pathogenicity
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0072700

Cell lines derived from the small intestine that reflect authentic properties of the originating intestinal epithelium are of high value for studies on mucosal immunology and host microbial homeostasis. A novel immortalization procedure was applied to generate continuously proliferating cell lines from murine E19 embryonic small intestinal tissue. The obtained cell lines form a tight and polarized epithelial cell layer, display characteristic tight junction, microvilli and surface protein expression and generate increasing transepithelial electrical resistance during in vitro culture. Significant up-regulation of Cxcl2 and Cxcl5 chemokine expression upon exposure to defined microbial innate immune stimuli and endogenous cytokines is observed. Cell lines were also generated from a transgenic interferon reporter (Mx2-Luciferase) mouse, allowing reporter technology-based quantification of the cellular response to type I and III interferon. Thus, the newly created cell lines mimic properties of the natural epithelium and can be used for diverse studies including testing of the absorption of drug candidates. The reproducibility of the method to create such cell lines from wild type and transgenic mice provides a new tool to study molecular and cellular processes of the epithelial barrier.