Human donor eyes, consented for research, were obtained from the Eye hospital and were approved by the Ethics Committee of Optometry Hospital affiliated to Wenzhou Medical University (2022-121-K-92-01). iPSCs were passaged onto Matrigel-coated dish in retinal differentiation medium (RDM, DMEM/F12 with 1% B27 (Gibco, 12587010), 1% N2 (Gibco, 17502048) and 1×NEAA (Gibco, 11140050). 10 mM nicotinamide (Selleck, S1899, Houston, Tx, USA), 10 ng/ml IGF1 (Sino Biological, 10598-HNAE, Beijing, China), 10 ng/ml bFGF (Sino Biological, 10014-HNAE), 1 M A-83-01 (Selleck, S7692) and 1 M IWR-1 (Selleck, S7086) were added to RDM for 0-4 days. 10 mM nicotinamide and 10 ng/ml bFGF were replaced by 100 ng/ml Activin A (Sino Biological, 10429-HNAH) and the concentration of IGF1 increased to 20 ng/ml, and A-83-01 was not added for 4-6 days. IGF1 and IWR-1 were replaced by 1 M SU5402 (Selleck, S7667) for 6-8 days. 1 M CHIR99021 (Selleck, S1263) was added to RDM for 8-14 days. From days 14, cells were maintained in RPE supporting medium (RSM, DMEM/F12 with 15% knockout serum replacement, 5% FBS, 1×NEAA and 10 mM nicotinamide) until they matured and formed pigmented monolayers. iPSC-RPE cells were passaged with EDTA (Gibco, 25200072) and were replaced on the Matrigel-coated dish and at P3 were used in this study. Total RNA was extracted using KAPA RNA prep kits (Roche, KK8560，Basel, Switzerland) from iPSC-RPE, hRPE and iPSC cells. Total RNA was subjected to ribosome removal, DNase digestion, fragmentation, and PCR amplification. Then libraries were sequenced on an Illuminar NovaSeq 6000.