PurposeTo develop a portable MR perfusion phantom for quality‐controlled assessment and reproducibility of arterial spin labeled (ASL) perfusion measurement.MethodsA 3D‐printed perfusion phantom was developed that mimics the branching of arterial vessels, capillaries, and a chamber containing cellulose sponge representing tissue characteristics. A peristaltic pump circulated distilled water through the phantom, and was first evaluated at 300, 400, and 500 mL/min. Longitudinal reproducibility of perfusion was performed using 2D pseudo‐continuous ASL at 20 post‐label delays (PLDs, ranging between 0.2 and 7.8 s at 0.4‐s intervals) over a period of 16 weeks, with three repetitions each week. Multi‐PLD data were fitted into a general kinetic model for perfusion quantification (f) and arterial transit time (ATT). Intraclass correlation coefficient was used to assess intersession reproducibility.ResultsMR perfusion signals acquired in the 3D‐printed perfusion phantom agreed well with the experimental conditions, with progressively increasing signal intensities and decreasing ATT for pump flow rates from 300 to 500 mL/min. The perfusion signal at 400 mL/min and the general kinetic model–derived f and ATT maps were similar across all PLDs for both intrasession and intersession reproducibility. Across all 48 experimental time points, the average f was 75.55 ± 3.83 × 10−3 mL/mL/s, the corresponding ATT was 2.10 ± 0.20 s, and the T1 was 1.84 ± 0.102 s. Intraclass correlation coefficient was 0.92 (95% confidence interval 0.83–0.97) for f, 0.96 (0.91–0.99) for ATT, and 0.94 (0.88–0.98) for T1, demonstrating excellent reproducibility.ConclusionA simple, portable 3D‐printed perfusion phantom with excellent reproducibility of 2D pseudo‐continuous ASL measurements was demonstrated that can serve for quality‐controlled and reliable measurements of ASL perfusion.