Abstract
AbstractPurpose Human pluripotent stem cell (hPSC) therapy is a potential approach for the replacement of degenerated retinal pigment epithelium (RPE) cells. For therapeutic use, safety and functionality of the RPE cells needs to be guaranteed. In addition to the basic cell and molecular biological characterizations it is vitally important to assess the differentiation status of acquired cells with functional tests.Methods We have previously shown that hPSC differentiate into RPE in xeno‐free and defined conditions. Recent advances as a culture of hPSC‐derived RPE cells sheets on clinically accepted material have led us to investigate the functionality of the RPE epithelium. The tightness of the epithelium was evaluated by measuring transepithelial electrical resistance and cell permeability. The phagocytic properties of hPSC‐RPE cells were studied using rat retinal explants.Results Human PSC‐derived putative RPE cells exhibits typical pigmented cobblestone‐like morphology and express RPE specific markers at both mRNA and protein level. In addition, cultured cells form a polarized epithelium with high integrity. In addition, co‐culturing hPSC‐RPE monolayers with rat retinal explants demonstrated that rhodopsin is internalized by cells in vitro.Conclusion We have demonstrated that hPSC‐RPE monolayers acquired RPE‐like properties, including characteristic RPE phenotype, expression of RPE markers and barrier functions. In addition, cells were capable of binding and internalizing rat rhodopsin when co‐cultured with rat retinal explants. Currently we investigate the correlations of the functions in the Royal College of Surgeons (RCS) rats.
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