Serial culture of murine primary airway epithelial cells and ex vivo replication of human rhinoviruses
Brockman-Schneider RA, Amineva SP, Bulat MV, Gern JE.
Department of Pediatrics and Medicine, University of Wisconsin-Madison, Madison
United States
Journal of Immunological Methods
Human rhinoviruses (HRV) are the primary etiological agents in cold infections, and represent a serious risk to individuals with chronic respiratory disease such as asthma. In order to develop treatment options for HRV infections, murine models are a crucial component in the study of infection mechanisms due to the wide array of reagents and techniques available to study murine immunology. We present here a cell culture system for studying isolated murine epithelial cell responses to HRV. Monolayers of primary mouse airway epithelial cells were maintained in a serial culture system, and the identity and purity of the cell population was confirmed via immunostaining (positive for cytokeratin, negative for vimentin). Infection of these cells with a minor group rhinovirus (HRV-1A) was evidenced by increases in viral RNA, de novo synthesis of viral proteins, and production of infectious virus. This model will be useful in experiments to define mechanisms of viral replication and host/virus interactions within airway epithelial cells.
Product use
Isolation and cultivation for immunostaining, infection with rhinovirus (HRV-1A), western blot and real-time PCR for detection of viral RNA replication. "„Submerged culture of harvested cells was then performed with a panel of various serum-free media and
Tissue type
Tissue info
Trachea from female, 7.5 week old C57BL/6N mice
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