title: Primary epithelial cell models for cystic fibrosis research.
abstract: 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 are 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.
authors.affiliation: Cystic Fibrosis/Pulmonary Research and Treatment Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. randell@med.unc.edu
authors.forename: Scott H
authors.initials: SH
authors.lastname: Randell
authors.forename: M Leslie
authors.initials: ML
authors.lastname: Fulcher
authors.forename: Wanda
authors.initials: W
authors.lastname: O'Neal
authors.forename: John C
authors.initials: JC
authors.lastname: Olsen
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D015925:Cryopreservation; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology* / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection
chemical_list: D003470:Culture Media; D007202:Indicators and Reagents; D034741:RNA, Small Interfering; D019005:Cystic Fibrosis Transmembrane Conductance Regulator