Abstract
Purpose :
Dry Eye Disease (DED) has a high prevalence of 5-50 % worldwide and thus impairs the quality of life of many individuals (Stapleton et al., Ocul. Surf. 2017; Jul(3)). Understanding the biology of the human tear film formation could help develop new drugs for DED treatment. We developed a 3D in vitro co-culture model of conjunctival and meibomian cells with the aim to increase predictivity of drug testing and for the reduction of animal experiments.
Methods :
Primary human conjunctival cells were isolated from healthy patient biopsies. Conjunctival fibroblasts were embedded in a collagen I matrix. Conjunctival epithelial cells (hCjE) were seeded on the apical side of the matrix and cultured at the air-liquid interface (Schwebler et al., Scientific Reports 2023, 13). Immortalized human meibomian gland epithelial cells (IHMGEC) were cultured in 80 % Matrigel to form organoids (adapted from Nuwormegbe et al., Ocul. Surf. 2022, Oct). After 2 weeks of culture, organoids were resuspended in 80 % Matrigel and placed on the periphery of the conjunctiva model. The co-culture model remained in culture for 15 days. Organoids and the co-culture model were analyzed via optical coherence tomography (OCT) and histology. Specific tissue markers were analyzed via immunostaining.
Results :
Meibomian organoids grew in size up to 100-200 µm while being cultured in proliferation medium. OCT showed the development of a multi-layered epithelium by hCjE. Furthermore, OCT revealed embedded organoids in the periphery of the model. Histological stainings confirmed a multi-layered conjunctival epithelium, including goblet cell differentiation as well as organoid structures of meibomian cells in the periphery of the model.
Conclusions :
We developed a novel co-culture model of conjunctival and meibomian cells that builds the base for studies on conjunctival and meibomian tissue and their interaction in vitro. In the future, drugs will be tested on this model to investigate effects on mucus or lipid secretion.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.