3D Oral and Cervical Tissue Models for Studying Papillomavirus Host-Pathogen Interactions

Area of Science:

• Biotechnology and Biomedical Engineering
• Cell Biology and Tissue Engineering

Background:

• Human papillomavirus (HPV) infection targets differentiating epithelial tissues, leading to cancers in oropharyngeal and anogenital sites.
• The HPV life cycle is intrinsically linked to epithelial cell differentiation, necessitating advanced tissue culture models.

Purpose of the Study:

• To develop and present two distinct, complementary methods for culturing three-dimensional (3D) stratified epithelial tissues.
• To create models that accurately recapitulate in vivo morphological and biochemical characteristics of human oral and cervical epithelia.
• To enable downstream applications for studying epithelial biology, HPV infection, and host-pathogen interactions.

Main Methods:

• Organotypic raft culture: Primary human epithelial cells grown on a collagen-fibroblast dermal equivalent at the liquid-air interface to promote stratification.
• Rotating wall vessel (RWV) bioreactor: Cells self-assemble into 3D aggregates on collagen-coated microbeads in a low-shear microgravity environment.
• Application to HPV-positive and HPV-negative oral and cervical cells, followed by various analytical techniques.

Main Results:

• Successful establishment of two distinct 3D epithelial tissue culture systems.
• Demonstration of critical morphological and biochemical mimicry of in vivo tissues.
• Application of these models to HPV-infected and uninfected oral and cervical cells for comprehensive analysis.

Conclusions:

• The developed methods provide robust 3D tissue models for studying HPV infection and epithelial biology.
• These 3D cultures are suitable for microscopic, ultrastructural, and omics-based investigations.
• The protocols facilitate research into HPV pathogenesis and host-pathogen interactions within relevant tissue contexts.