Electrical stimulation has demonstrated great effectiveness in the modulation of cell fate in vitro and regeneration therapy in vivo. Conventionally, the employment of electrical signal comes with the electrodes, battery, and connectors in an invasive fashion. This tedious procedure and possible infection hinder the translation of electrical stimulation technologies in regenerative therapy. Given electromechanical coupling and flexibility, piezoelectric polymers can overcome these limitations as they can serve as a self-powered stimulator via scavenging mechanical force from the organism and external stimuli wirelessly. Wireless electrical cue mediated by electrospun piezoelectric polymeric nanofibers constitutes a promising paradigm allowing the generation of localized electrical stimulation both in a noninvasive manner and at cell level. Recently, numerous studies based on electrospun piezoelectric nanofibers have been carried out in electrically regenerative therapy. In this review, brief introduction of piezoelectric polymer and electrospinning technology is elucidated first. Afterward, we highlight the activating strategies (e.g., cell traction, physiological activity, and ultrasound) of piezoelectric stimulation and the interaction of piezoelectric cue with nonelectrically/electrically excitable cells in regeneration medicine. Then, quantitative comparison of the electrical stimulation effects using various activating strategies on specific cell behavior and various cell types is outlined. Followingly, this review explores the present challenges in electrospun nanofiber-based piezoelectric stimulation for regeneration therapy and summarizes the methodologies which may be contributed to future efforts in this field for the reality of this technology in the clinical scene. In the end, a summary of this review and future perspectives toward electrospun nanofiber-based piezoelectric stimulation in tissue regeneration are elucidated.