Propagation of infectious, high-risk HPV in organotypic "raft" culture.

Margaret E. McLaughlin-Drubin, Craig Meyers

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)171-186
Number of pages16
JournalMethods in molecular medicine
Volume119
StatePublished - 2005

All Science Journal Classification (ASJC) codes

  • Molecular Medicine

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