From: Long-Term Efficacy of GMP Grade Xeno-Free hESC-Derived RPE Cells Following Transplantation Trans. Vis. Sci. Tech.. 2017;6(3):17. doi:10.1167/tvst.6.3.17 Figure Legend: Characterization of OpRegen 5B cells. (A) FACS analysis demonstrating that 99.01% of OpRegen 5B cells coexpress the RPE markers CRALBP and PMEL17. (B) Immunostaining of cultured OpRegen 5B cells followed by confocal imaging and manual counting of 500 DAPI-positive cells using the ImageJ software, demonstrated that 95% of the cells in the culture were positive for Bestrophin 1 and 90% positive for MITF. (C) OpRegen 5B cells cultured on a transwell for 5 weeks demonstrated their ability to generate barrier function (TEER > 100 Ω) as well as secrete PEDF and VEGF in a polarized manner, PEDF to the apical side (apical to basal PEDF ratio > 1) and VEGF to the basal side (basal to apical VEGF ratio > 1). Data represent average ± standard deviation (SD) of four TEER-, PEDF-, and VEGF-independent measurements of OpRegen five subbatches (5A–5D that were cryopreserved over 4 days). (D) Using a highly sensitive qualified TRA-1-60/Oct4 double staining FACS method (LOD 0.0004%; LOQ [limit of quantification] 0.001%; accuracy and precision ≤ 25%, linearity R2 > 0.99 in the range 0.001%–1%), no hESCs (levels below assay LOD) were detected in OpRegen 5B cells (10 × 106 cells were acquired). Date of download: 10/24/2017 The Association for Research in Vision and Ophthalmology Copyright © 2017. All rights reserved.