Purpose: To investigate the effect of acquisition parameters on the accuracy of 2D velocity encoded cine magnetic resonance imaging (VEC MRI) flow measurements. Materials and Methods: Using a pulsatile flow phantom, through-plane flow measurements were performed on a flexible vessel made of polyvinyl alcohol cryogel (PVA), a material that mimics the MR signal and biomechanical properties of aortic tissue. Results: Repeated VEC MRI flow measurements (N = 20) under baseline conditions yielded an error of 0.8 +/- 1.5%. Slice thickness, angle between flow and velocity encoding directions, spatial resolution, velocity encoding range, and radio frequency (RF) flip angles were varied over a clinically relevant range. Spatial resolution had the greatest impact on accuracy, with a 9% overestimation of flow at 16 pixels per vessel cross-section. Conclusion: VEC MRI proved to be an accurate and reproducible technique for pulsatile flow measurements over the range of acquisition parameters examined as long as sufficient spatial resolution was prescribed.