Non-thermal plasmas have recently been applied to biomedical challenges, including blood coagulation, sterilization and malignant cell apoptosis. We previously demonstrated that non-thermal dielectric barrier discharge plasma promotes angiogenesis through reactive oxygen species-induced fibroblast growth factor-2 release. However, it remained unclear which plasma-produced reactive species contributed to this effect. In this study, H2O2, OH•, O, and O2(1Δg) were measured in liquid and cells after plasma treatment using specific fluorescent probes. While all reactive species except O2(1Δg) increased with plasma dose in both liquid and cells, H2O2 and OH• were primarily responsible for plasma-induced angiogenesis. This study will guide development of new plasma devices for angiogenesis in wound healing and tissue engineering.