As the economic challenges associated with vision impairment and ocular injuries continue to escalate, there exists an urgent need to explore innovative remedies for these conditions. Extracellular Vesicles (EVs), containing a diverse array of biological components such as nucleic acids, proteins, and lipids, have garnered substantial scientific support for their healing and immune-regulating attributes in addressing various disorders. Nevertheless, an area that remains relatively uncharted is the potential reparative impact of stem cell-derived EVs within the realm of ocular damage or disease. Furthermore, prevailing literature predominantly focuses on EVs derived from primary stem cells, neglecting a comprehensive assessment of EVs derived from hTERT-immortalized stem cells. Through our extensive EV production platform, we have consistently generated EVs sourced from hTERT mesenchymal stem cells (MSCs) and have employed diverse techniques to characterize and profile their cargo contents. Leveraging established cellular assays, we have conducted a comparative analysis of these EVs' effects on both intact and impaired retinal pigment epithelial cells. To the best of our knowledge, this marks the inaugural study demonstrating the feasibility of consistent, large-scale production of hTERT MSC EVs and exploring their potential regenerative properties against compromised ocular cells. The outcomes of our investigations affirm that hTERT MSCs elicit multiple reparative influences in vitro, underscoring their promise not only as a viable substitute for primary MSC-derived EVs but also as a prospective innovative therapeutic agent for addressing ocular injuries.