Spiro-OMeTAD hole transport materials typically exhibit an amorphous state in perovskite solar cells. However, the lack of structural ordering leads to weak intermolecular interaction, inferior carrier transfer, and poor stability in devices. Herein, we developed a π-conjugation-induced short-range ordering strategy to modulate the stacking order of spiro-OMeTAD during film formation. A clear molecular ordering at the nanoscale is observed, which enhances intermolecular π-π stacking in spiro-OMeTAD and enables effective carrier extraction and favorable energy level alignment. The nanoscale-ordered spiro-OMeTAD allows the achievement of perovskite solar cells with a champion efficiency of 25.37%, surpassing devices utilizing amorphous spiro-OMeTAD (23.52%). The unencapsulated device demonstrates enhanced operational stability by retaining 98% of its initial efficiency under continuous 1 sun equivalent illumination at 60 °C for 840 h. This work establishes a significant and valid modulation concept for the stacking order of organic transport materials, paving the way for the development of efficient and stable perovskite solar cells.