Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Bacteria / immunology
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Bacteria / pathogenicity
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Biomarkers / metabolism
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Cell Line
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Cell Polarity / physiology
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Cell Proliferation
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Embryo, Mammalian
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Immunity, Innate / physiology*
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Interferon Type I / metabolism
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Intestinal Mucosa / cytology*
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Intestinal Mucosa / embryology
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Intestinal Mucosa / immunology*
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Intestine, Small / cytology*
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Intestine, Small / embryology
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Intestine, Small / immunology*
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Mice
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Primary Cell Culture / methods*
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Viruses / immunology
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Viruses / pathogenicity
Substances
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Biomarkers
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Interferon Type I
Grants and funding
This study was supported by funding from the Deutsche Forschungsgemeinschaft for the Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy), the research grant Ho 2236/8-1, the priority program SPP1656 and the Collaborative Research Center SFB900 (Chronic Infection), the German Ministry of Education and Research (01KI1003D) and an Europäischer Fonds für regionale Entwicklung grant (80029155) from the “Niedersächsisches Ministerium für Wissenschaft und Kultur”. JS was supported by the Hannover Biomedical Research School, the Center for Infection Biology, as well as by the Helmholtz Centre for Infection Research GradSchool. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.