Full thickness 3D in vitro conjunctiva model enables goblet cell differentiation

Scientific Reports · 4 authors, 3 centres

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TL;DR

Researchers developed a 3D in vitro model of human conjunctiva by combining epithelial cells with a fibroblast-embedded collagen hydrogel. This full-thickness model successfully produced a non-keratinized, multi-layered epithelium containing mucus-secreting goblet cells, unlike simpler epithelial-only models.

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The study developed a full-thickness (FTConM) and a simple epithelial (rhConE) 3D in vitro model using primary human conjunctival epithelial cells and fibroblasts. The combination of epithelial cells with a connective tissue equivalent (fibroblasts in a collagen hydrogel) resulted in a non-keratinized stratified epithelium with goblet cell differentiation, evidenced by cup-like morphology, Alcian blue/PAS staining, and MUC5AC expression. Key factors for goblet cell differentiation were identified as the presence of conjunctival fibroblasts, the collagen matrix, and specific medium supplements. The model closely resembles natural conjunctiva and provides a platform for studying conjunctival diseases and drug testing.

STUDY TYPE

other

SPECIALTY

ophthalmology

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ORIGINAL PAPER

PMC10382544

Full thickness 3D in vitro conjunctiva model enables goblet cell differentiation

Scientific Reports

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ABSTRACT

In vitro culture and generation of highly specialized goblet cells is still a major challenge in conjunctival 3D in vitro equivalents. A model comprising all physiological factors, including mucus-secreting goblet cells has the potential to act as a new platform for studies on conjunctival diseases. We isolated primary conjunctival epithelial cells and fibroblasts from human biopsies. 3D models were generated from either epithelial layers or a combination of those with a connective tissue equivalent. Epithelial models were investigated for marker expression and barrier function. Full-thickness models were analyzed for goblet cell morphology and marker expression via immunofluorescence and quantitative real-time PCR. Simple epithelial models cultured at the air–liquid interface showed stratified multi-layer epithelia with pathologic keratinization and without goblet cell formation. The combination with a connective tissue equivalent to generate a full-thickness model led to the formation of a non-keratinized stratified multi-layer epithelium and induced goblet cell differentiation. In our model, a high resemblance to natural conjunctiva was achieved by the combination of conjunctival epithelial cells with fibroblasts embedded in a collagen-hydrogel as connective tissue equivalent. In the future, our conjunctival in vitro equivalent enables the investigation of goblet cell differentiation, conjunctival pathologies as well as drug testing.

CITATION EXPORT

Julian Schwebler, Christina Fey, Daniel Kampik et al.. Full thickness 3D in vitro conjunctiva model enables goblet cell differentiation. Scientific Reports. (2023). https://doi.org/10.1038/s41598-023-38927-8

AUTHORS

4 · 3 CENTRES

Julian Schwebler, Christina Fey, Daniel Kampik, Christian Lotz

1grid.424644.40000 0004 0495 360XTranslational Center Regenerative Therapies (TLC-RT), Fraunhofer Institute for Silicate Research (ISC), Würzburg, Germany
2grid.411760.50000 0001 1378 7891Chair of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg, Würzburg, Germany
3grid.411760.50000 0001 1378 7891Department of Ophthalmology, University Hospital Würzburg, Würzburg, Germany