Background Advances in sub-Nyquist-sampled dynamic contrast-enhanced (DCE) MRI enable monitoring of brain tumors with millimeter resolution and whole-brain coverage. Such undersampled quantitative methods need careful characterization regarding achievable test-retest reproducibility. Purpose To demonstrate a fully automated high-resolution whole-brain DCE MRI pipeline with 30-fold sparse undersampling and estimate its reproducibility on the basis of reference regions of stable tissue types during multiple posttreatment time points by using longitudinal clinical images of high-grade glioma. Materials and Methods Two methods for sub-Nyquist-sampled DCE MRI were extended with automatic estimation of vascular input functions. Continuously acquired three-dimensional k-space data with ramped-up flip angles were partitioned to yield high-resolution, whole-brain tracer kinetic parameter maps with matched precontrast-agent T1 and M0 maps. Reproducibility was estimated in a retrospective study in participants with high-grade glioma, who underwent three consecutive standard-of-care examinations between December 2016 and April 2019. Coefficients of variation and reproducibility coefficients were reported for histogram statistics of the tracer kinetic parameters plasma volume fraction and volume transfer constant (Ktrans) on five healthy tissue types. Results The images from 13 participants (mean age ± standard deviation, 61 years ± 10; nine women) with high-grade glioma were evaluated. In healthy tissues, the protocol achieved a coefficient of variation less than 57% for median Ktrans, if Ktrans was estimated consecutively. The maximum reproducibility coefficient for median Ktrans was estimated to be at 0.06 min-1 for large or low-enhancing tissues and to be as high as 0.48 min-1 in smaller or strongly enhancing tissues. Conclusion A fully automated, sparsely sampled DCE MRI reconstruction with patient-specific vascular input function offered high spatial and temporal resolution and whole-brain coverage; in healthy tissues, the protocol estimated median volume transfer constant with maximum reproducibility coefficient of 0.06 min-1 in large, low-enhancing tissue regions and maximum reproducibility coefficient of less than 0.48 min-1 in smaller or more strongly enhancing tissue regions. Published under a CC BY 4.0 license. Online supplemental material is available for this article. See also the editorial by Lenkinski in this issue.