title: Generation and Infection of Organotypic Cultures with Epstein-Barr Virus.
abstract: While numerous model systems are available to study EBV latency in B cells and have contributed greatly to our understanding of the role of these cells in the viral life cycle, models to study the EBV life cycle in epithelial cells in vitro are lacking. Epithelial cells are poorly infected in vitro, and EBV-infected cell lines have not been successfully obtained from epithelial tumors. Recently, we have demonstrated that organotypic cultures of oral keratinocytes can be used as a model to study EBV infection in the epithelial tissue. These "raft" cultures generate a stratified tissue resembling the epithelium seen in vivo with a proliferating basal layer and differentiating suprabasal layers. Here, we describe generation of EBV-infected raft cultures established from primary oral mucosal epithelial cells, which exhibit high levels of productive replication induced by differentiation, as well as methods to analyze EBV infection.
authors.affiliation: Department of Microbiology and Immunology, H107, The Pennsylvania State University College of Medicine, 500 University Drive, Hershey, PA, 17033, USA.|Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, 03755, USA.
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Temple
authors.affiliation: Department of Microbiology and Immunology, H107, The Pennsylvania State University College of Medicine, 500 University Drive, Hershey, PA, 17033, USA.|The Penn State Hershey Cancer Institute, Hershey, PA, 17033, USA.
authors.forename: Craig
authors.initials: C
authors.lastname: Meyers
authors.affiliation: Department of Microbiology and Immunology, H107, The Pennsylvania State University College of Medicine, 500 University Drive, Hershey, PA, 17033, USA. csample@hmc.psu.edu.|The Penn State Hershey Cancer Institute, Hershey, PA, 17033, USA. csample@hmc.psu.edu.
authors.forename: Clare E
authors.initials: CE
authors.lastname: Sample
mesh_terms: D000818:Animals; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology; D020031:Epstein-Barr Virus Infections / Q000821:virology*; D016679:Genome, Viral; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000821:virology; D051379:Mice; D009061:Mouth Mucosa / Q000378:metabolism / Q000473:pathology / Q000821:virology; D009924:Organ Culture Techniques; D060888:Real-Time Polymerase Chain Reaction; D019562:Viral Load; D014779:Virus Replication
keywords: Epithelial cells; Epstein–Barr virus; Lytic replication; Organotypic culture; Stratified epithelium; Virus production; qPCR