Despite the recent progress in the field of conductive polymers, a deeper understanding is still required to explain the differences between reported data. In this work, we reveal that the electrical conductivity σ of a poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) film can be significantly increased by spin-coating multiple thin layers onto a substrate. Generally, σ can be improved by more than fourfold for multiple layers, as compared to a single thicker one. A gradual enhancement is observed for pristine PEDOT:PSS films (up to 2.10±0.26 S cm −1 for 5 layered films), while a plateau in σ at around 200 S cm −1 is reached after only three layers, when using a PEDOT:PSS solution with 5 vol.% dimethyl sulfoxide (DMSO). In contrast, only a small change in σ is observed for single layers of varying thickness. Accordingly, the thermoelectric power factor is also increased by up to 3.4 times for the multiple layers. Based on AFM, XPS, UV-Vis and Raman spectroscopy measurements, two mechanisms are also proposed, involving an increase in percolation by inclusion of smaller grains within the existing ones, respectively a reorganization of the PEDOT:PSS chains. Firstly, these findings represent a direct strategy for enhancing the thermoelectric performance of conductive polymer films without additional reagents. Secondly, the mechanistic insights contribute to understand the broader picture, by explaining existing literature results.