Methods in Molecular Biology 2011-01-01

Primary epithelial cell models for cystic fibrosis research.

Scott H Randell, M Leslie Fulcher, Wanda O'Neal, John C Olsen

Index: Methods Mol. Biol. 742 , 285-310, (2011)

Full Text: HTML

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.