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Derivation of Traceable and Transplantable Photoreceptors from Mouse Embryonic Stem Cells

2014; Elsevier BV; Volume: 2; Issue: 6 Linguagem: Inglês

10.1016/j.stemcr.2014.04.010

2213-6711

Sarah Decembrini, Ute Koch, Freddy Radtke, Alexandre Moulin, Yvan Arsenijévic,

bioluminescence and chemiluminescence research

Retinal 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.(N-NIII) Qualitative colocalization analysis of Manders' coefficient visualized on the images as white spots.(NI) mGluR6 versus PKC-a, (NII) GFP versus PKC-a, and (NIII) GFP versus mGluR6 channel are shown.DAPI nuclear stain is in blue.**p < 0.01 and ***p < 0.001 by ANOVA with Tukey's correction.P4, postnatal day 4 retina-derived photoreceptors (n = 4); DA, dark adapted; LA, light adapted.Scale bars, 10 mm (A, D, E, and F), 4 mm (CI, DI, EI, FI, G, and H), 15 mm (J), 5 mm (inset in J), 5 mm (N), and 1 mm (inset in NI).