A pumpless fuel supply using pressurized fuel with autonomous flow regulation valves is presented. Since micropumps and their control circuitry consume a portion of the electrical power generated in fuel cells, fuel supply without micropumps makes it possible to provide more efficient and inexpensive fuel cells than conventional ones. The flow regulation valves in the present system maintain the constant fuel flow rate from the pressurized fuel chamber even though the fuel pressure decreases. They autonomously adjust fluidic resistance of the channel according to fuel pressure so as to maintain constant flow rate. Compared to previous pumpless fuel supply methods, the present method offers more uniform fuel flow without any fluctuation using a simple structure. The prototypes were fabricated by a polymer micromolding process. In the experimental study using the pressurized deionized water, prototypes with pressure regulation valves showed constant flow rate of 5.38 ± 0.52 μl/s over 80 min and 5.89 ± 0.62 μl/s over 134 min, for the initial pressure in the fuel chamber of 50 and 100 kPa, respectively, while the other prototypes having the same fluidic geometry without flow regulation valves showed higher and gradually decreasing flow rate. The present pumpless fuel supply method providing constant flow rate with autonomous valve operation will be beneficial for the development of next-generation fuel cells.