在实验室培养的原代人鼻上皮细胞中诱导粘膜屏障保护炎症反应

Current protocols in toxicology Pub Date : 2019-02-04 DOI:10.1002/cptx.69

Mahnaz Ramezanpour, Harrison Bolt, Alkis Psaltis, Peter-John Wormald, Sarah Vreugde

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引用次数: 12

摘要

在这里，我们使用toll样受体(TLR) 3激动剂poly I:C (LMW)在人鼻上皮细胞(HNECs)的浸没和/或气液界面(ALI)培养细胞中诱导炎症反应。通过酶联免疫吸附试验(ELISA)测定白细胞介素-6 (IL-6)蛋白水平来确定炎症反应。粘膜屏障的完整性是通过测量经上皮电阻(TEER)和荧光标记右旋糖酐的传代来确定的。poly (I:C) LMW刺激诱导HNEC-ALI细胞产生15- 17倍的IL-6。©2019 by John Wiley &儿子,Inc。

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Inducing a Mucosal Barrier–Sparing Inflammatory Response in Laboratory-Grown Primary Human Nasal Epithelial Cells

Here we use the toll-like receptor (TLR) 3 agonist poly I:C (LMW) to induce an inflammatory response in cells of submerged and/or air-liquid interface (ALI) cultures of human nasal epithelial cells (HNECs). The inflammatory response is determined by measuring interleukin-6 (IL-6) protein levels by enzyme-linked immunosorbent assay (ELISA). The mucosal barrier integrity is determined by measuring transepithelial electrical resistance (TEER) and passage of fluorescently labeled dextrans. Stimulation with poly (I:C) LMW induces a 15- to 17-fold increase in IL-6 production by HNEC-ALI cells. © 2019 by John Wiley & Sons, Inc.

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Current protocols in toxicology

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