Nine Latin American countries plan to use silvopastoral practices—incorporating trees into grazing lands—to mitigate climate change. However, the cumulative potential of scaling up silvopastoral systems at national levels is not well quantified. Here, we combined previously published tree cover data based on 250 m resolution MODIS satellite remote sensing imagery for 2000–2017 with ecofloristic zone carbon stock estimates to calculate historical and potential future tree biomass carbon storage in Colombian grasslands. Between 2000 and 2017, tree cover across all Colombian grasslands increased from 15% to 18%, with total biomass carbon (TBC) stocks increasing from 0.41 to 0.48 Pg. The range in 2017 carbon stock values in grasslands based on ecofloristic zones (5 to 122 Mg ha⁻¹) suggests a potential for further increase. Increasing all carbon stocks to the current median and 75th percentile levels for the respective eco-floristic zone would increase TBC stocks by about 0.06 and 0.15 Pg, respectively. Incorporated into national C accounting, such Tier 2 estimates can set realistic targets for silvopastoral systems in nationally determined contributions (NDCs) and nationally appropriate mitigation actions (NAMAs) implementation plans in Colombia and other Latin American countries with similar contexts.