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Culturing and Applications of Rotating Wall Vessel Bioreactor Derived 3D Epithelial Cell Models
Published on: April 3, 2012
Development and characterization of a three-dimensional organotypic human vaginal epithelial cell model
Brooke E Hjelm1, Alice N Berta, Cheryl A Nickerson
1Center for Infectious Diseases and Vaccinology, The Biodesign Institute at Arizona State University, 1001 South McAllister Avenue, Tempe, AZ 85287-5401, USA.
Biology of Reproduction
|December 17, 2009
Summary
A novel 3-D human vaginal epithelial cell model was created using rotating wall vessel bioreactor technology. This advanced model accurately mimics in vivo conditions, offering a superior platform for testing microbicide safety and efficacy against sexually transmitted infections.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Microbiology
Background:
- Current in vitro models often fail to fully replicate the complex in vivo human vaginal environment.
- Existing methods for microbicide testing, such as animal models or explants, have limitations in throughput and human relevance.
- Developing a more physiologically relevant in vitro model is crucial for accurate assessment of microbicide candidates.
Purpose of the Study:
- To develop and characterize a novel three-dimensional (3-D) in vitro human vaginal epithelial cell (EC) model.
- To assess the utility of this 3-D model for high-throughput toxicity testing of microbicides.
- To compare the response of the 3-D model to a 2-D cell culture system when exposed to a known microbicide candidate.
Main Methods:
- Human vaginal ECs were cultured on microcarrier beads in a rotating wall vessel (RWV) bioreactor to form 3-D aggregates.
- Structural and functional properties were analyzed using immunofluorescence and electron microscopy.
- The model's response to toll-like receptor (TLR) agonists and nonoxynol-9 (N-9) was evaluated by quantifying cytokine production and inflammatory biomarkers.
Main Results:
- The RWV-generated 3-D vaginal EC model successfully recapitulated key in vivo features, including stratification, microvilli, tight junctions, and mucus production.
- The 3-D model demonstrated functional expression of TLRs 2, 3, 5, and 6.
- The 3-D aggregates exhibited greater resistance to nonoxynol-9 (N-9) compared to 2-D monolayers, and N-9 treatment induced dose-dependent inflammatory biomarkers.
Conclusions:
- The developed 3-D vaginal EC model provides a robust and physiologically relevant platform for microbicide research.
- This model can serve as a valuable complementary tool for screening microbicide safety and efficacy, potentially improving clinical trial outcomes.
- The 3-D model's enhanced resistance to N-9 and induction of inflammatory markers offer critical insights into microbicide-host interactions.
