Fig. 3 hESC-RPE cell sheet transplantation and validation of the surgical method. hESC-RPE cell sheet transplantation and validation of the surgical method.

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Date of download: 6/2/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Spectral Domain Optical Coherence Tomography: Ultra-high.

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Fig. 3 hESC-RPE cell sheet transplantation and validation of the surgical method. hESC-RPE cell sheet transplantation and validation of the surgical method. (A) Steps for the preparation and loading of the hESC-RPE cell sheet on the hAM scaffold into the injection device. (B) Image of the hESC-RPE cell sheet embedded in gelatin. A small piece (2 to 3 mm2) was cut for injection into the subretinal space of the rat eye (black rectangle). (C) Image showing loading of the hESC-RPE cell sheet (black rectangle) into the head of the injection device. (D) OCT analysis of the retina of a wild-type control rat and a dystrophic RCS rat after transplantation of the hESC-RPE cell sheet (red line, dystrophic + graft). (E to G) Representative immunofluorescence confocal microscopic images of sections of retina from athymic nude rats 10 days after transplantation with either a hESC-RPE cell sheet (left) or a hESC-RPE cell suspension (right). Sections were stained for human MTCO2 and human ZO-1 (E), human MERTK and human MTCO2 (F), or human MERTK and human collagen IV (G). Nuclei were visualized with a DAPI counterstain. Three rat retinas transplanted with hESC-RPE cell suspensions and four rat retinas transplanted with hESC-RPE cell sheets were analyzed. White box indicates regions that are enlarged. Images correspond to maximal projections of z stacks. Scale bars, 50 μm and 10 μm (higher magnification). IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; OLM, outer limiting membrane. Karim Ben M’Barek et al., Sci Transl Med 2017;9:eaai7471 Published by AAAS