Human Papillomaviruses pp 171-186

Part of the Methods in Molecular Medicine book series (MIMM, volume 119)

Propagation of Infectious, High-Risk HPV in Organotypic “Raft” Culture

  • Margaret E. McLaughlin-Drubin
  • Craig Meyers


The organotypic (raft) culture system has been used to develop an in vitro system that is capable of reproducing the entire human papillomavirus (HPV) life cycle, including virion morphogenesis. This system utilizes HPV-containing cell lines that are either derived from biopsies or created by the transfection of keratinocytes with HPV genomic DNA. When grown as raft cultures, these lines allow for a detailed study of all stages of the viral life cycle. In this chapter, we describe in detail how to (1) culture keratinocytes, (2) electroporate primary keratinocytes with HPV DNA, (3) detect episomal HPV genomes by Southern (DNA) blotting, (4) grow organotypic raft cultures, (5) isolate HPV, and (6) perform in vitro infectivity testing.


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Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Margaret E. McLaughlin-Drubin
    • 1
  • Craig Meyers
    • 1
  1. 1.The Department of Microbiology and ImmunologyThe Pennsylvania State University College of MedicineHershey

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