Two-dimensional (2D) Ruddlesden-Popper (RP) perovskites have emerged as attractive candidates for high-performance perovskite solar cells (PSCs) thanks to their superior environmental and structural stability. However, 2D RP PSCs exhibit larger exciton binding energy due to the dielectric mismatch between the organic and inorganic layers, resulting in poorer photovoltaic performance compared to their 3D analogs. Here, we developed a selenophene-based spacer, namely, 2-selenophenemethylammonium (SeMA), for stable and efficient 2D RP PSCs. The 2D perovskite film using methylammonium (MA) as the A-site cation (nominal n = 5) shows excellent film quality with large grain size and a preferred vertical orientation relative to the substrate. Furthermore, we have successfully demonstrated the effectiveness of a predeposition transport layer (PDTL) consisting of [6,6]-phenyl-C61-butyric acid methyl ester (PCBM) in passivating surface defects of the perovskite film and inducing densification of the upper PCBM electron transport layer. This densification promotes efficient extraction and transport of electrons. The optimized PSCs based on 2D RP perovskite using MA as A-site cation (nominal n = 5) achieved a power conversion efficiency (PCE) of 17.25%, which was further boosted to 19.03% when using formamidinium (FA) as A-site cation. This represents a record PCE of 2D RP PSCs by using the selenophene-based spacer. Moreover, these 2D RP PSCs significantly improve thermal, moisture, and light stability. Our results provide significant implications for the synergistic strategy of developing selenophene-based spacers and device engineering methods for achieving highly efficient and stable 2D RP perovskite solar cells.