Abstract
Purpose :
Hereditary choroidopathies and Age-related macular degeneration (AMD) gradually lead to the death of retinal cells, resulting in irreversible loss of sight. Currently there are no effective treatments. Retinal cells are protected and nourished by the blood-retinal barrier (BRB). The BRB consists of the cells of the retinal pigment epithelium on one side and the choriocapillaries on the other with the Bruch membrane in between. The BRB is affected in different ways in these diseases. Our project aims to develop 3D cell culture models of retinal pigment epithelium (RPE)/choroidal cells derived from pluripotent stem cells using a biodegradable material with adequate physical-chemical and mechanical properties to mimic the BRB. In this context, three-dimensional co-culture represent very important tools to investigate the relationships between the two cell layers. They help in increasing our knowledge of these chorioretinal diseases and to find new treatments.
Methods :
In the preliminary experiments presented, we have assays for the establishment of 3-D cell culture using human amniotic membrane as the scaffold. To this end, we have cultured the hESC-derived RPE cells and the most commonly used human endothelial cells, the HUVEC cell line on an amniotic membrane.
Results :
Microscopy observations showed that the RPE cells formed a typical hexagonal cobblestone monolayer epithelium on the apical side of amniotic membrane while the HUVEC cells attached on the opposite face of the amniotic membrane surface.
Conclusions :
These preliminary results showed the possibility of the co-culture of the RPE cells and the HUVEC cell line in a 3D-culture system.
The next step will be to replace the amniotic membrane with a biodegradable material with adequate physical-chemical and mechanical properties to mimic the BRB.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.