Department of Gene Regulation and Differentiation, Helmholtz Centre for Infection Research, Braunschweig, Germany.
PLoS One. 2013 Aug 5;8(8):e72700. doi: 10.1371/journal.pone.0072700. Print 2013.
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.
从小肠衍生而来的细胞系反映了起源于肠道上皮的真实特性，对于粘膜免疫学和宿主微生物稳态的研究具有很高的价值。本研究应用一种新的永生化程序从小鼠 E19 胚胎小肠组织中生成可连续增殖的细胞系。所获得的细胞系形成紧密且极化的上皮细胞层，显示出特征性的紧密连接、微绒毛和表面蛋白表达，并在体外培养过程中产生逐渐增加的跨上皮电阻。暴露于特定的微生物先天免疫刺激物和内源性细胞因子时，观察到趋化因子 Cxcl2 和 Cxcl5 的表达显著上调。还从小鼠干扰素报告基因（Mx2-Luciferase）中转基因创建了细胞系，允许基于报告基因技术定量细胞对 I 型和 III 型干扰素的反应。因此，新创建的细胞系模拟了天然上皮的特性，可用于包括测试候选药物吸收在内的各种研究。该方法从野生型和转基因小鼠中创建此类细胞系的重现性为研究上皮屏障的分子和细胞过程提供了新工具。
