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. 2021 Feb;9(3):245. doi: 10.21037/atm-20-4707

Figure 1.

Figure 1

Maturation and characterization of human iRPE cells. (A) Scheme of GMP-compatible processes to manufacture and test clinical-grade iPSCs and iRPE cells for preclinical studies. (B) Timeline of manufacturing clinical-grade iRPE cells from iPSC WCB. A GMP-compatible immature iRPE master cell bank was established ~26 days from the start of monolayer iPSC differentiation, and all reagents used were xeno-free and chemically defined. (C) Gene expression analysis of known RPE-specific genes of iRPE cells collected on days 35 and 45 of differentiation. (D) Top panel: Bright field images of the iRPEs in monolayer on days 20 and 40 of differentiation. Bottom panel: Immunostaining of the mature iRPE (day 40) markers ZO-1 (red) and MiTF (green). Scale bars, 100 µm. (E) Flow cytometric analysis of mature iRPE cells (day 40) costained for MiTF/RPE65, MiTF/Best1, RPE65/ZO-1, MiTF/Otx2, and MiTF/ZO-1. WCB, working cell bank; RIM, RPE induction medium; RDM, RPE differentiation medium; RMM, RPE maturation medium.