Nine Latin American countries have identified silvopastoral practices—trees and livestock systems combined—as a key component of their Nationally Determined Contributions (Paris Climate Accord) or Nationally Appropriate Mitigation Actions (Bali Action Plan). However, the climate change mitigation potential of silvopastoral systems is not well studied. This study estimates the mitigation potential of silvopastoral practices via carbon sequestration in woody biomass in Colombian grassland. We derived tree cover maps from the MODIS Version 6 Vegetation Continuous Field MOD44B dataset for 2000-2017 and employed the Intergovernmental Panel on Climate Change Tier 1 default carbon storage estimates to calculate current and potential above- and below- ground biomass carbon stocks in existing grasslands. Total tree cover across all Colombian grasslands increased by about 20% from year 2000 (15%) to 2017 (18%). Concomitantly, the total (above- and belowground biomass carbon combined) biomass carbon (TBC) stocks increased by about 17% from 2000 (mean = 34 t C ha-1; SD =18) to 2017 (mean = 39 t C ha-1; SD =18). TBC stocks in Colombian grassland were 0.41 and 0.48 Petagrams (Pg) C in 2000 and 2017, respectively. Average 2017 carbon stock values ranged from 5 and 222 t C ha-1; this suggests potential for increasing carbon stocks in areas of low tree cover. About 73% of the total grassland had TBC less than the <75th percentile. Increasing all carbon stocks to the current median and 75th percentile levels would increase total carbon stocks by about 0.06 Pg C (0.2 Pg CO2e) and 0.15 Pg C (0.57 Pg CO2e), respectively. This study provides valuable data on the climate change mitigation potential of silvopastoral system as part of the Nationally Determined Contributions and Nationally Appropriate Mitigation Actions of Colombia and other Latin American countries. We highlighted on some of the potential impacts of increasing tree cover on grassland biomes. Limitations include a likely underestimation of tree cover, and an assumed direct linear relationship between tree cover and biomass were identified.