The versatility of conducting polymers (CPs) facilitates their use in energy conversion and storage, sensor, and biomedical technologies, once processed into thin films. Hydrophobic CPs, like poly(3,4-ethylenedioxythiophene) (PEDOT), typically require the use of surfactant additives, such as poly(styrenesulfonate) (PSS), to aid their aqueous processability as thin films. However, excess PSS diminishes CP electrochemical performance, biocompatibility, and device stability. Here, we report the electrosynthesis of PEDOT thin films at a polarised liquid|liquid interface, a method non-reliant on conductive solid substrates that produces free-standing, additive-free, biocompatible, easily transferrable, and scalable 2D PEDOT thin films of any shape or size in a single-step at ambient conditions. We demonstrate the PEDOT thin film’s superior biocompatibility as scaffolds for cellular growth, opening immediate applications in organic electrochemical transistor (OECT) devices for monitoring cell behaviour over extended time periods, bio-scaffolds and medical devices, without the requirement for physiologically unstable and poorly biocompatible PSS.