Several series and parallel Blumlein-type striplines designed on multilayered printed circuit board (PCB) are connected to match different impedance loads with different pulsewidths to develop a high-voltage nanosecond pulse generator for investigating the biological effects of a nanosecond pulsed electric field. First, the principle of a multistage series-parallel Blumlein stripline system is described. In particular, the multilayered strip transmission line is designed on the five-layered PCB to minimize the volume of the system whose maximum size is 300 mm (length) × 260 mm (width) × 154 mm (height), and the gate resistor capacitor and diode active equalizing circuit is used as the dynamic/static balancing tactic. Then, a singlestage Blumlein-type stripline and some multistage series-parallel Blumlein-type stripline are established to test their performance. As a result, the nanosecond pulse generator produces a rectangular pulse on a matched resistor load, with an amplitude of 0-1.7 kV, a pulsewidth of integer multiples of 100 ns, a rise time of ∼20 ns or less, and a repetition frequency of 0 to several kHz. A variety of biomedical loads can be matched by parallel stacking mode, which will facilitate the analysis of the biological effects of the nanosecond pulsed electric field.