title: Using Organotypic Epithelial Tissue Culture to Study the Human Papillomavirus Life Cycle.
abstract: Human papillomaviruses (HPVs) are small double-stranded DNA viruses that are associated with greater than 95% of cervical cancers and 20% of head and neck cancers. These cancers arise from persistent infections in which there is continued expression of the HPV E6 and E7 oncogenes, often as a consequence of integration of HPV DNA into the host genome. Such cancers represent "dead ends" for the virus as integration disrupts the viral genome and because the cancers are defective in normal epithelial differentiation, which is required for production of progeny papillomavirus. In order to study the full viral life cycle, from the establishment to maintenance to productive stages, our lab makes use of the organotypic epithelial tissue culture system. This system allows us to mimic the three-dimensional structure of epithelia whose differentiation is tightly linked to the completion of the HPV viral life cycle. In this chapter we describe how various aspects of the HPV life cycle are monitored in raft cultures making use of an immortalized keratinocyte cell line. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
authors.forename: Denis
authors.initials: D
authors.lastname: Lee
authors.affiliation: Flow Cytometry Laboratory, Carbone Cancer Center, University of Wisconsin, Madison, Wisconsin.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Norby
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
authors.forename: Mitchell
authors.initials: M
authors.lastname: Hayes
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
authors.forename: Ya-Fang
authors.initials: YF
authors.lastname: Chiu
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
authors.forename: Bill
authors.initials: B
authors.lastname: Sugden
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
authors.forename: Paul F
authors.initials: PF
authors.lastname: Lambert
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D004848:Epithelium / Q000821:virology*; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000821:virology*; D014776:Virus Cultivation / Q000379:methods*
keywords: keratinocyte; life cycle; organotypic epithelial tissue culture; papillomavirus