Recapitulation of microvascular structure, function and perfusion in vitro can enable studies of vascular biology, provide a model for diseases such as ischemic stroke or tumor angiogenesis and enable quantitative evaluation of physiologic blood or lymph perfusion. Here we describe the initial design and deployment of a first-generation, self-contained 3D-printed, physiologically-faithful, microfluidic perfusion phantom to form explicit, hierarchically-branching, microvascular structure encapsulated in a type I collagen matrix in vitro, with pump-driven perfusion easily visible via phase-contrast MRI (Fig. 1). The phantom flexibly supports creation of user-defined vessel network geometries with human vascular cells and allows experimental validation of blood flow, i.e., via constitutive equations for convective and diffusive transport that quantitatively relate the flux of tracers from time-resolved images to transport field quantities. Thus, the largely qualitative and unmeasurable global arterial input assumption in the traditional Kety’s method can be replaced with measurable and reproducible MRI experimental data, formulated as quantitative transport mapping (QTM). Preliminary data demonstrate that the QTM phantom is promising for characterizing actual blood transport in vitro in healthy and pathological contexts.