Despite the great potential of polymer microfibers in human-friendly wearable electronics, most previous polymeric electronics have been limited to thin-film-based devices due to practical difficulties in fabricating microfibrillar devices, as well as defining the active channel dimensions in a reproducible manner. Herein, we report on conducting polymer microfiber-based organic electrochemical transistors (OECTs) and their application in single-strand fiber-type wearable ion concentration sensors. We developed a simple wet-spinning process to form very conductive poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) microfibers using aqueous sulfuric acid solutions and carefully examined their electrical/electrochemical properties. In conjunction with fabricating substrate-free PEDOT:PSS microfiber-based OECT devices, the proposed novel characterization method demonstrated that the current variation ratio can be a reliable method for evaluating the device performance for sensing ion concentrations, regardless of the actual channel dimensions. Finally, we developed single-strand fiber-type skin-mountable OECTs by introducing a source-gate hybrid electrode and demonstrated that the resultant microfiber sensors can perform real-time repetitive measurements of the ion concentration in human sweat.