Derivation of Traceable and Transplantable Photoreceptors from Mouse Embryonic Stem Cells

•De novo isolation of Crx-GFP embryonic stem cell lines to trace photoreceptors•3D culture system fine-tuning to generate many integration-competent photoreceptors•Revealing in-vitro- and in-vivo-developing retina similarities•Characterization of the most appropriate stage to transplant photoreceptors

SummaryRetinal degenerative diseases resulting in the loss of photoreceptors are one of the major causes of blindness. Photoreceptor replacement therapy is a promising treatment because the transplantation of retina-derived photoreceptors can be applied now to different murine retinopathies to restore visual function. To have an unlimited source of photoreceptors, we derived a transgenic embryonic stem cell (ESC) line in which the Crx-GFP transgene is expressed in photoreceptors and assessed the capacity of a 3D culture protocol to produce integration-competent photoreceptors. This culture system allows the production of a large number of photoreceptors recapitulating the in vivo development. After transplantation, integrated cells showed the typical morphology of mature rods bearing external segments and ribbon synapses. We conclude that a 3D protocol coupled with ESCs provides a safe and renewable source of photoreceptors displaying a development and transplantation competence comparable to photoreceptors from age-matched retinas.