Groundwater is an increasingly important water supply source globally. Understanding the amount of groundwater used versus the volume available is crucial to evaluate future water availability. We present a groundwater stress assessment to quantify the relationship between groundwater use and availability in the world's 37 largest aquifer systems. We quantify stress according to a ratio of groundwater use to availability, which we call the Renewable Groundwater Stress ratio. The impact of quantifying groundwater use based on nationally reported groundwater withdrawal statistics is compared to a novel approach to quantify use based on remote sensing observations from the Gravity Recovery and Climate Experiment (GRACE) satellite mission.