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Optimized protocol for 3D epithelial cultures supporting human papillomavirus replication.

Marta Laganà1, Gabriela Cuesta Margolles1, Agnieszka Jaracz-Ros1

  • 1INSERM UMR-996, Inserm, Inflammation, Microbiome and Immunosurveillance, Faculté de Pharmacie, Université Paris-Saclay, 91400 Orsay, France.

STAR Protocols
|January 21, 2024
PubMed
Summary

This study presents an updated 3D full-thickness epithelial culture (3Dep) method to support the complete human papillomavirus (HPV) life cycle. This optimized protocol facilitates research into HPV pathogenesis and replication in keratinocytes.

Keywords:
Cell BiologyCell cultureMicrobiologyOrganoids

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Area of Science:

  • Virology
  • Cell Biology
  • Tissue Engineering

Background:

  • Human papillomaviruses (HPVs) are common viruses with disease-causing potential.
  • HPV replication depends on keratinocyte (KC) proliferation and differentiation.
  • Existing methods have limitations for studying the complete HPV life cycle.

Purpose of the Study:

  • To provide an updated, optimized workflow for generating 3D full-thickness epithelial cultures (3Deps).
  • To enable the complete HPV replicative life cycle in a 3D epithelial model.
  • To facilitate research on HPV infection and pathogenesis.

Main Methods:

  • Adaptation of the organotypic epithelial raft culture protocol.
  • Detailed steps for HPV genome preparation and KC transfection.
  • Procedures for dermal equivalent preparation, 3Deps culture, and analysis.

Main Results:

  • Successfully generated pluristratified human epithelia supporting HPV replication.
  • Established a robust 3D model for studying HPV life cycle dynamics.
  • Provided a comprehensive workflow for HPV research.

Conclusions:

  • The updated 3Dep culture system is effective for studying the complete HPV life cycle.
  • This model offers a valuable tool for investigating HPV pathogenesis and developing antiviral strategies.
  • The protocol is amenable to various HPV types and research applications.