Self-assembly of organic sensitizer layers in cobalt complex-based DSCs was studied to elucidate its role in reducing the loss of charge recombination. DSCs with various dye loadings were fabricated by dye desorption without the aid of basic solvent. The FT-IR and UV results indicate the deprotonation of the anchoring organic sensitizers, which influences the conduction band of TiO2 remarkably by changing the surface potential. Positive band edge shifts and a decrease of the recombination rate constant are demonstrated to be the main factors affecting energy loss at open circuit. In contrast, absorbed photon conversion efficiency (APCE) analyses illuminate the crucial role of the packing of the anchoring sensitizer in reducing recombination loss at short circuit. This is further supported by numerical simulations, which show that APCE is primarily dependent on the recombination rate constant rather than the band edge shift at short circuit. These results highlight the importance of self-assembly of sensitizers with insulating groups in retarding charge recombination by forming overlapping molecular layers.