Primary Epithelial Cell Models for Cystic Fibrosis Research
Primary Epithelial Cell Models for Cystic Fibrosis Research
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DOI：
10.1007/978-1-61779-120-8_18
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Olsen, John C.
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文献类型：
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作者：
Randell, Scott H.;Fulcher, M. Leslie;Olsen, John C.
当原代人气道上皮 (hAE) 细胞在气液界面 (ALI) 的多孔支撑物上体外生长时，它们会重现体内形态和关键生理过程。这些培养物可用于研究呼吸道生物学和疾病以及测试新的囊性纤维化 (CF) 疗法。本章提供了使用非专有试剂创建分化良好的原代 CF 和非 CF 气道上皮细胞培养物的方案。我们还讨论了逆转录病毒和慢病毒载体的生产、hAE 细胞系的衍生、报告基因测定以及 ALI hAE 培养物中基因过度表达和敲低的不断发展的科学。
When primary human airway epithelial (hAE) cells are grown in vitro on porous supports at an air-liquid interface (ALI), they recapitulate in vivo morphology and key physiologic processes. These cultures arc useful for studying respiratory tract biology and diseases and for testing new cystic fibrosis (CF) therapies. This chapter gives protocols enabling creation of well-differentiated primary CF and non-CF airway epithelial cell cultures with non-proprietary reagents. We also discuss the production of retroviral and lentiviral vectors, the derivation of hAE cell lines, reporter gene assays, and the evolving science of gene overexpression and knockdown in ALI hAE cultures.