In pursuit of authenticity: IPS cell-derived RPE for clinical applications

Authors

• K Miyagishima
• Q Wan
• B Corneo
• R Sharma
• M Lotfi
• N Boles
• F Hua
• A Maminishkis
• Congxiao
• TM Blenkinsop
• V Khristov
• B Jha
• O Memon
• S D’Souza
• S Temple
• S Miller
• K Bharti

Abstract

The RPE is a monolayer of pigmented, hexagonal cells connected by tight junctions that support photoreceptor function and maintain health/integrity of the distal retina and adjacent choroidal blood supply. iPSCs can be efficiently differentiated into retinal pigment epithelial cells (RPE), offering the possibility of autologous cell replacement therapy for retinal degeneration stemming from RPE loss. The generation and maintenance of epithelial apical/basolateral polarity is fundamental for iPSC- derived RPE (iPSC- RPE) to recapitulate native RPE structure/function. Presently, no criteria have been established to determine clonal or donor based heterogeneity in the polarization and maturation state of iPSC- RPE. Here we provide an unbiased structural, molecular, and physiological evaluation of 15 iPSC- RPE derived from distinct tissues in several different donors. We assessed the intact RPE monolayer in terms of an ATP- dependent signaling pathway that drives critical aspects of RPE function, including calcium and electrophysiological responses, as well as steady - state fluid transport. These responses have key in vivo counterparts that together help determine the homeostasis of the distal retina. In particular, the measurement of fluid flow from the apical to basal side of the RPE monolayer is a sensitive and informative assay to authenticate iPSC-RPE polarization and function. We characterized the donor and clonal variation and found that iPSC- RPE function was more significantly affected by the genetic differences between different donors than the epigenetic differences associated with different starting tissues. This study provides a reference dataset to authenticate genetically diverse iPSC- RPE derived for clinical applications.

This page was last updated on Friday, March 26, 2021