The retina is a complex, multi-layered tissue responsible for sensing and converting light photons to neural signals for visual recognition. The outermost cells of the tissue, the retinal pigment epithelium (RPE), performs a number of critical functions in the visual cycle, and is subjected to enormous daily stress to meet these various demands. Over time and particularly in disease, the RPE undergoes a number of well-characterised changes, however the molecular mechanisms involved in these changes are not completely understood. Using single-cell sequencing technology, we aim to assess the unique transcriptional changes that occur in biologically aged human pluripotent stem cell-derived RPE cells. The human embryonic stem cell (hESC) line H9 was differentiated to RPE using an adaptation of our published protocol (Lidgerwood et al 2016). Briefly, cells were differentiated for 60 days, passaged and maintained continuously until the characteristic pigmented, polygonal and homogeneous morphology was observed at day 30 (D30) post-passaging, at which point the first sample, designated “young” RPE, was harvested. The remaining culture was continuously maintained and harvested on day 365 (D365), designated “aged” RPE. Both samples at the time of harvesting were live cell sorted using FACS and captured on the 10X Genomics Single Cell 3′ Chip. Sequencing libraries were generated with unique sample indices (SI) for each sample and mapped to the Homo sapien genome (GRCh38, Annotation: Gencode v29). Using standard processing and analysis, we identified 652 uniquely differentially expressed genes between the young vs aged hPSC-RPE, which were largely involved in response to metal ions, oxidation, trafficking, immune responses and mitochondrial function, according to (GO) biological processes. Together this data is the first to map the biological transcriptome of aged hPSC-RPE at the single cell level and may provide insight into the early molecular events that lead to RPE aging and age-related diseases.