scholarly journals Characterization of Human Coronaviruses on Well-Differentiated Human Airway Epithelial Cell Cultures

2015 ◽  
pp. 73-87 ◽  
Author(s):  
Hulda R. Jonsdottir ◽  
Ronald Dijkman
Keyword(s):  
Epithelial Cell ◽  
Cell Cultures ◽  
Airway Epithelial Cell ◽  
Airway Epithelial ◽  
Epithelial Cell Cultures ◽  
Human Airway ◽  
Human Airway Epithelial Cell ◽  
Well Differentiated ◽  
Human Coronaviruses

Well-Differentiated Human Airway Epithelial Cell Cultures

2004 ◽  
pp. 183-206 ◽  
Author(s):  
M. Leslie Fulcher ◽  
Sherif Gabriel ◽  
Kimberlie A. Burns ◽  
James R. Yankaskas ◽  
Scott H. Randell
Keyword(s):  
Epithelial Cell ◽  
Cell Cultures ◽  
Airway Epithelial Cell ◽  
Airway Epithelial ◽  
Epithelial Cell Cultures ◽  
Human Airway ◽  
Human Airway Epithelial Cell ◽  
Well Differentiated

scholarly journals Establishment of Primary Transgenic Human Airway Epithelial Cell Cultures to Study Respiratory Virus–Host Interactions

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 747 ◽  
Author(s):  
Hulda R. Jonsdottir ◽  
Sabrina Marti ◽  
Dirk Geerts ◽  
Regulo Rodriguez ◽  
Volker Thiel ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Genetic Modification ◽  
Airway Epithelial Cell ◽  
Respiratory Viruses ◽  
Target Cells ◽  
Airway Epithelial ◽  
Host Interactions ◽  
Human Airway ◽  
Human Airway Epithelial Cell

Primary human airway epithelial cell (hAEC) cultures represent a universal platform to propagate respiratory viruses and characterize their host interactions in authentic target cells. To further elucidate specific interactions between human respiratory viruses and important host factors in the airway epithelium, it is important to make hAEC cultures amenable to genetic modification. However, the short and finite lifespan of primary cells in cell culture creates a bottleneck for the genetic modification of these cultures. In the current study, we show that the incorporation of the Rho-associated protein kinase (ROCK) inhibitor (Y-27632) during cell propagation extends the life span of primary human cells in vitro and thereby facilitates the incorporation of lentivirus-based expression systems. Using fluorescent reporters for fluorescence-activated cell sorting (FACS)-based sorting, we generated homogenously fluorescent hAEC cultures that differentiate normally after lentiviral transduction. As a proof-of-principle, we demonstrate that host gene expression can be modulated post-differentiation via inducible short hairpin (sh)RNA-mediated knockdown. Importantly, functional characterization of these transgenic hAEC cultures with exogenous poly (I:C), as a proxy for virus infection, demonstrates that such modifications do not influence the host innate immune response. Moreover, the propagation kinetics of both human coronavirus 229E (HCoV-229E) and human respiratory syncytial virus (hRSV) were not affected. Combined, these results validate our newly established protocol for the genetic modification of hAEC cultures, thereby unlocking a unique potential for detailed molecular characterization of virus–host interactions in human respiratory epithelium.

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