The objective of the research study was to determine the ability of Lactobacillus rhamnosus ATCC 9595, Lactobacillus casei ATCC 334 and strains of lactic acid bacteria isolated from faeces of healthy children to bind cholesterol in the environment of model intestinal juice and MRS Broth medium. A cholesterol RTU enzymatic kit was used to determine the cholesterol residue in samples. The cultures were grown in 24-well titration plates. In order to determine the degree of cholesterol binding and OD600, the absorbance was measured using a Spark 10M microplate reader. The strains with the highest degree of cholesterol binding were evaluated for their ability to grow and bind cholesterol from the environment with the increasing cholesterol load (1.66 g/dm³, 2.85 g/dm³, 3.75 g/dm³). L. rhamnosus and L. casei were able to bind 68.18 % and 83.6 % of cholesterol from the MRS Broth medium, respectively, while in the model intestinal juice those values were 60 % and 50 %, respectively. The strains isolated from the faeces of children were characterised by a high variability as regards the degree of binding cholesterol from the environment, whereas better results were achieved for the model intestinal juice (1.81 ÷ 77.99 %) than for the MRS Broth medium (0.49 ÷ 33.14 %). With the increasing amount of cholesterol in the environment, the strains tested showed a reduced increase in the biomass and this was correlated with the reduced degree of binding of that compound. As regards the model intestinal juice, the highest reduction in the  level of cholesterol contained therein was obtained at its concentration of 1.66 g/dm³ . The cholesterol  concentration of 3.75 g/dm³ caused the increase in the biomass to be inhibited and thus the lowest quantity of this compound was removed from the environment.