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In Vitro Method to Study Sex-Based Differences in Conjunctival Goblet Cells
Published on: July 28, 2023
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Full thickness 3D in vitro conjunctiva model enables goblet cell differentiation.
Julian Schwebler1,2, Christina Fey1, Daniel Kampik2
1Translational Center Regenerative Therapies (TLC-RT), Fraunhofer Institute for Silicate Research (ISC), Würzburg, Germany.
Scientific Reports
|July 28, 2023
Summary
Creating a 3D conjunctival in vitro equivalent model using fibroblasts and epithelial cells successfully induced goblet cell differentiation. This advanced model mimics natural conjunctiva for studying diseases and drug testing.
Area of Science:
- Ophthalmology
- Tissue Engineering
- Cell Biology
Background:
- Generating specialized goblet cells in conjunctival 3D in vitro equivalents remains a significant challenge.
- A comprehensive model including mucus-secreting goblet cells is crucial for studying conjunctival diseases.
Purpose of the Study:
- To develop a functional 3D conjunctival in vitro equivalent that incorporates mucus-secreting goblet cells.
- To establish a novel platform for investigating conjunctival pathologies and evaluating drug efficacy.
Main Methods:
- Isolation of primary human conjunctival epithelial cells and fibroblasts.
- Generation of 3D models using epithelial layers alone or combined with a connective tissue equivalent (fibroblasts in collagen-hydrogel).
- Analysis of cell morphology, marker expression (immunofluorescence), barrier function, and gene expression (qRT-PCR).
Main Results:
- Epithelial models cultured at the air-liquid interface formed stratified epithelia but lacked goblet cells and showed keratinization.
- Full-thickness models incorporating a connective tissue equivalent induced non-keratinized stratified epithelia and goblet cell differentiation.
- The developed model closely resembles natural conjunctiva.
Conclusions:
- Combining conjunctival epithelial cells with a fibroblast-based connective tissue equivalent is essential for inducing goblet cell differentiation in 3D in vitro models.
- This novel conjunctival in vitro equivalent serves as a valuable tool for research on goblet cell biology, conjunctival diseases, and therapeutic interventions.
