Differentiation of human bronchial epithelial cells (HBEs) in air-liquid interface (ALI) cultures recapitulates organotypic modeling of the in vivo environment. Although ALI cultures are invaluable for studying the respiratory epithelial barrier, loss-of-function studies are limited by potentially cytotoxic reagents in classical transfection methods, the length of the differentiation protocol, and the number of primary epithelial cell passages. Here, we present the efficacy and utility of a simple method for siRNA transfection of HBEs in ALI cultures that does not require potentially cytotoxic transfection reagents and does not detrimentally alter the physiology of HBEs during the differentiation process. This transfection protocol introduces a reproducible and efficient method for loss-of-function studies in HBE ALI cultures that can be leveraged for modeling the respiratory system and airway diseases.

中文翻译：

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气液界面气道上皮细胞培养中基因敲低的被动 siRNA 转染方法

空气-液体界面 (ALI) 培养物中人支气管上皮细胞 (HBE) 的分化概括了体内环境的器官模型。尽管 ALI 培养物对于研究呼吸道上皮屏障非常宝贵，但功能丧失研究受到经典转染方法中潜在的细胞毒性试剂、分化方案的长度和原代上皮细胞通道数的限制。在这里，我们介绍了一种在 ALI 培养物中对 HBE 进行 siRNA 转染的简单方法的功效和实用性，该方法不需要潜在的细胞毒性转染试剂，并且在分化过程中不会有害地改变 HBE 的生理。