Grassland ecosystems play an important role in subsistence agriculture and the global carbon cycle. However, the global spatio-temporal patterns and environmental controls of grassland biomass are not well quantified and understood. The goal of this study was to estimate the spatial and temporal patterns of the global grassland biomass and analyze their driving forces using field measurements, Normalized Difference Vegetation Index (NDVI) time series from satellite data, climate reanalysis data, and a satellite-based statistical model. Results showed that the NDVI-based biomass carbon model developed from this study explained 60% of the variance across 38 sites globally. The global carbon stock in grassland aboveground live biomass was 1.05 Pg·C, averaged from 1982 to 2006, and increased at a rate of 2.43 Tg·C·y−1 during this period. Temporal change of the global biomass was significantly and positively correlated with temperature and precipitation. The distribution of biomass carbon density followed the precipitation gradient. The dynamics of regional grassland biomass showed various trends largely determined by regional climate variability, disturbances, and management practices (such as grazing for meat production). The methods and results from this study can be used to monitor the dynamics of grassland aboveground biomass and evaluate grassland susceptibility to climate variability and change, disturbances, and management.