Abstract
Purpose :
Retinitis pigmentosa (RP) and age-related macular degeneration (AMD) are the main causes of blindness in the developed world. Stem cell-based therapy represents an alternative approach in the treatment of retinal diseases. The Retinal Pigment Epithelium (RPE) is a continuous monolayer of cuboidal epithelial cells, localized between the photoreceptors and fenestrated choroid capillaries. The RPE interacts with the photoreceptors for the maintenance of visual function. RP and AMD could be caused by degeneration or malfunction of the RPE cell layer. Two important features of human embryonic stem cells (hESC), self-renew and the ability to generate an unlimited number of RPE cells, make them very attractive tools in the cell therapy approach for retinal diseases. Maintaining the epithelial morphology of RPE cells is a crucial parameter to consider in order restoring some visual function by cell therapy.
Methods :
To achieve this goal, we have developed a tissue engineered product (TEP) that consists of RPE derived from hESC cultured on denuded human amniotic membrane (hAM). In our study, we have optimized a differentiation protocol allowing us to generate RPE from hESC in a large amount using xeno-free culture media.
Results :
Our results show that more than 98% of differentiated cells expressed RPE typical markers. In parallel, we have established high quality standard to definite their characteristics and impurities. This cells could be banked and maintain their phenotype when cultured on hAM. In fact, when cultured for 1 month on denuded hAM, the hESC-derived RPE cells formed a typical hexagonal pigmented epithelium. RPE cells secrete VEGF in polarized manner (basal >apical).
Conclusions :
Finally, our TEP will be used for phase I/II clinical trials for the treatment of RP caused by a RPE defect.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.