Molecular profiling of stem cell-derived retinal pigment epithelial cell differentiation established for clinical translation

Sandra Petrus-Reurer, Alex R Lederer, Laura Baqué-Vidal, Iyadh Douagi, Belinda Pannagel, Irina Khven, Monica Aronsson, Hammurabi Bartuma, Magdalena Wagner, Andreas Wrona, Paschalis Efstathopoulos, Elham Jaberi, Hanni Willenbrock, Yutaka Shimizu, J Carlos Villaescusa, Helder André, Erik Sundstrӧm, Aparna Bhaduri, Arnold Kriegstein, Anders Kvanta, Gioele La Manno, Fredrik Lanner

Abstract

Human embryonic stem cell-derived retinal pigment epithelial cells (hESC-RPE) are a promising cell source to treat age-related macular degeneration (AMD). Despite several ongoing clinical studies, a detailed mapping of transient cellular states during in vitro differentiation has not been performed. Here, we conduct single-cell transcriptomic profiling of an hESC-RPE differentiation protocol that has been developed for clinical use. Differentiation progressed through a culture diversification recapitulating early embryonic development, whereby cells rapidly acquired a rostral embryo patterning signature before converging toward the RPE lineage. At intermediate steps, we identified and examined the potency of an NCAM1+ retinal progenitor population and showed the ability of the protocol to suppress non-RPE fates. We demonstrated that the method produces a pure RPE pool capable of maturing further【24†source】.