About: Establishment of Primary Transgenic Human Airway Epithelial Cell Cultures to Study Respiratory Virus

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About: Establishment of Primary Transgenic Human Airway Epithelial Cell Cultures to Study Respiratory Virus–Host Interactions Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Establishment of Primary Transgenic Human Airway Epithelial Cell Cultures to Study Respiratory Virus–Host Interactions
Creator	Thiel, Volker Dijkman, Ronald Jonsdottir, Hulda Rodriguez, Regulo Geerts, Dirk Marti, Sabrina
Source	Medline; PMC
abstract	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.
has issue date	2019-08-13(xsd:dateTime)
bibo:doi	10.3390/v11080747
bibo:pmid	31412613
has license	cc-by
sha1sum (hex)	f442fd7cf9f7ab6a2939acae3d1eea548dafac20
schema:url	https://doi.org/10.3390/v11080747
resource representing a document's title	Establishment of Primary Transgenic Human Airway Epithelial Cell Cultures to Study Respiratory Virus–Host Interactions
has PubMed Central identifier	PMC6723040
has PubMed identifier	31412613
schema:publication	Viruses
resource representing a document's body	covid:f442fd7cf9f7ab6a2939acae3d1eea548dafac20#body_text
is schema:about of	named entity 'HUMAN' named entity 'fluorescent' named entity 'specific' named entity 'factors' named entity 'Y-27632' named entity 'Epithelial' named entity 'TRANSGENIC' named entity 'RESULTS' named entity 'IN VITRO' named entity 'PRIMARY CELLS' named entity 'CHARACTERIZATION' named entity 'PLATFORM' named entity 'NEWLY' named entity 'DIFFERENTIATION' named entity 'HUMAN CELLS' named entity '28I' named entity 'TARGET CELLS' named entity 'CULTURES' named entity 'POST' named entity 'SYSTEMS' named entity 'INDUCIBLE' named entity 'SHORT' named entity 'CURRENT' named entity '229E' named entity 'PROTOCOL' named entity 'MOLECULAR' named entity 'EXPRESSION' named entity 'GENE EXPRESSION' named entity 'MODULATED' named entity 'GENETIC MODIFICATION' named entity 'ESTABLISHED' named entity 'STUDY' named entity 'PROXY' named entity 'AIRWAY EPITHELIUM' named entity 'BASED' named entity 'MEDIATED' named entity 'PROOF' named entity 'HUMAN RESPIRATORY SYNCYTIAL VIRUS' named entity 'FLUORESCENCE-ACTIVATED CELL SORTING' named entity 'VIRUS INFECTION' named entity 'OUR' named entity 'PRINCIPLE' named entity 'REPORTERS' named entity 'IMPORTANT' named entity 'TRANSDUCTION' named entity 'PRIMARY' named entity 'RESPIRATORY EPITHELIUM' named entity 'DETAILED' named entity 'LENTIVIRUS' named entity 'FLUORESCENT' named entity 'AFFECTED' named entity 'INFLUENCE' named entity 'PROPAGATION' named entity 'HOST' named entity 'RESPIRATORY VIRUS' named entity 'STUDY' named entity 'INTERACTIONS' named entity 'AIRWAY EPITHELIAL CELL' named entity 'OF-' named entity 'COMBINED' named entity 'HOST' named entity 'HUMAN' named entity 'DIFFERENTIATE' named entity 'SORTING' named entity 'THEIR' named entity 'LIFESPAN' named entity 'THESE' named entity 'TRANSGENIC' named entity 'EXOGENOUS' named entity 'MAKE' named entity 'CELL' named entity 'KINETICS' named entity 'POLY' named entity 'INTERACTIONS' named entity 'AIRWAY EPITHELIAL CELL' named entity 'EXTENDS'

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