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Structurally and functionally characterized in vitro model of rabbit vocal fold epithelium
Masanobu Mizuta1, Takashi Kurita2, Emily E Kimball3
1Department of Otolaryngology-Head and Neck Surgery, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawaharacho, Sakyo-ku, Kyoto 606-8507, Japan.
Tissue & Cell
|April 12, 2017
Summary
This study establishes a rabbit vocal fold epithelial cell culture model to assess epithelial barrier function. Optimized culture conditions, including seeding density and passage number, yield a well-differentiated cell multilayer for TEER measurements.
Area of Science:
- Cell Biology
- Tissue Engineering
- Otorhinolaryngology
Background:
- The vocal fold epithelium serves as a critical barrier.
- In vitro models are needed to study vocal fold epithelial barrier function.
- Understanding barrier integrity is crucial for vocal fold health.
Purpose of the Study:
- To develop a method for primary culture of rabbit vocal fold epithelial cells.
- To establish a reliable in vitro model for evaluating epithelial barrier function using transepithelial electrical resistance (TEER).
- To optimize culture conditions for a well-differentiated epithelial cell multilayer.
Main Methods:
- Rabbit larynges were enzymatically treated to isolate vocal fold epithelial cells for primary culture.
- Cells were co-cultured with feeder cells on collagen-coated plates and passaged.
- Transepithelial electrical resistance (TEER) was measured to assess barrier function under various conditions (growth additives, seeding density, passage number).
- Epithelial differentiation and tight junction protein expression were confirmed via histology and immunostaining.
Main Results:
- Fetal bovine serum supplementation resulted in higher TEER compared to bovine pituitary extract.
- Optimal seeding density for TEER was determined to be 2.2×10^4 cells/cm^2.
- Passage two exhibited higher TEER than passage three, indicating better barrier function.
- Immunostaining confirmed the expression of epithelial markers (CK13, CK14) and tight junction proteins (occludin, ZO-1).
Conclusions:
- A robust in vitro model for rabbit vocal fold epithelial cells was successfully developed.
- The optimized culture method yields a well-differentiated epithelial multilayer with functional tight junctions.
- This model provides a valuable tool for investigating epithelial barrier function and potential therapeutic interventions for vocal fold disorders.