Abstract
Extracellular vesicles (EVs) are a class of naturally occurring secreted cellular bodies that are involved in long distance cell-to-cell communication. Proteins, lipids, mRNA, and miRNA can be packaged into these vesicles and released from the cell. This information is then delivered to target cells. Since EVs are naturally adapted molecular messengers, they have emerged as an innovative, inexpensive, and robust method to deliver therapeutic cargo in vitro and in vivo. Well-differentiated primary cultures of human airway epithelial cells (HAE) are refractory to standard transfection techniques. Indeed, common strategies used to overexpress or knockdown gene expression in immortalized cell lines simply have no detectable effect in HAE. Here we use EVs to efficiently deliver siRNA or protein to HAE. Furthermore, EVs can deliver CFTR protein to cystic fibrosis donor cells and functionally correct the Cl− channel defect in vitro. EV-mediated delivery of siRNA or proteins to HAE provides a powerful genetic tool in a model system that closely recapitulates the in vivo airways.
Highlights
Primary cultures of well-differentiated human airway epithelial cells (HAE) are a robust model for studying epithelial cell biology
HAE prepared from donor trachea and bronchi are grown at an air–liquid interface and form a pseudostratified columnar epithelium with tight junctions
The cultures include multiple cell types such as ciliated cells, non-ciliated cells, goblet cells, and basal cells that recapitulate the surface cells of the conducting airways [2,46]. These cells provide an excellent model for studying airway cell biology
Summary
Primary cultures of well-differentiated human airway epithelial cells (HAE) are a robust model for studying epithelial cell biology. Cells grown at an air–liquid interface form a polarized, pseudostratified columnar epithelium that closely resembles the morphology of the in vivo surface epithelium of the conducting airways [1,2,3]. This model provides an opportunity to study cell biology, disease progression, pathogenesis, and treatments for lung diseases like cystic fibrosis [4]. Transfecting siRNA into poorly differentiated airway cells leads to knockdown of target genes; this strategy has its limitations because poorly differentiated HAE are less representative of the in vivo airways. Viral vectors can be expensive and time consuming to generate
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