Primary open-angle glaucoma (POAG) is an optic neuropathy characterized by gradual degeneration of retinal ganglion cells (RGCs) leading to irreversible vision loss and, if untreated, blindness. Upon diagnosis treatment options are limited and often fail to halt the progression of vision loss. This is due to the incomplete understanding of the disease. The molecular profiling of human RGCs in normal and diseased tissue is hindered by the lack of non-invasive means for obtaining RGCs from living donors. This can now be circumvented by the use of induced pluripotent stem cells (iPSCs) as a source of RGCs. With the use of an automated platform, we generated 305 human iPSC lines from glaucoma patients (n = 143) and healthy controls (n = 162). All lines underwent quality control analysis consisting of virtual karyotyping and assessment of pluripotency markers OCT-4 and TRA-1-60.Subsequently, 183 lines were differentiated into RGCs via retinal organoids and then subjected to single cell RNA sequencing (scRNA-seq) to gain in-depth information about transcriptomic differences between healthy controls and glaucoma patient samples. Understanding mechanisms underlying RGC function, maintenance of homeostasis and those conferring susceptibility to POAG is crucial to discover new therapeutic targets and commence the process of drug discovery.