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

ABSTRACT: Human papillomaviruses (HPVs) are commensal viruses with pathogenic potential. Their life cycle requires the proliferation and differentiation of keratinocytes (KCs) to form pluristratified epithelia. Based on the original organotypic epithelial raft cultures protocol, we provide an updated workflow to optimally generate pluristratified human epithelia supporting the complete HPV replicative life cycle, here called 3D full-thickness epithelial cultures (3Deps). We describe steps for HPV genome preparation, KC transfection, and dermal equivalent preparation. We then detail procedures for 3Deps culture, harvesting, and analysis.

SUBMITTER: Lagana M

PROVIDER: S-EPMC10835287 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

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

Laganà Marta M Margolles Gabriela Cuesta GC Jaracz-Ros Agnieszka A Mercier-Nomé Françoise F Roingeard Philippe P Lambert Paul F PF Schlecht-Louf Géraldine G Bachelerie Françoise F

STAR protocols 20240119 1

Human papillomaviruses (HPVs) are commensal viruses with pathogenic potential. Their life cycle requires the proliferation and differentiation of keratinocytes (KCs) to form pluristratified epithelia. Based on the original organotypic epithelial raft cultures protocol, we provide an updated workflow to optimally generate pluristratified human epithelia supporting the complete HPV replicative life cycle, here called 3D full-thickness epithelial cultures (3Deps). We describe steps for HPV genome p ...[more]

PMID: 38245871

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