Stingless bees (Apidae: Meliponini) are culturally and economically important bees and they represent a major component of the Colombian melittofauna. However, the bee fauna of Colombia is still in early stages of exploration and species identification is often difficult or impossible. We revised the species of the cleptobiotic stingless bee genus Lestrimelitta in Colombia and recognized the following eleven species: L. glabrata, L. guyanensis, L. rufa, L. rufipes, and L. spinosa, which are recorded for the first time for the country; L. huilensis, L. opita, L. piedemontana, and two new species, L. diminuta, sp. n. and L. galvisi sp. n. We also confirmed the presence of L. limao in the Colombian Amazon and provide new geographical records with an updated key to species from Central America and northern South America