This work evaluated the abilities of five isolates of lactic acid bacteria (LAB) from different sources, i.e Lactobacillus fermentum S21209 and Lactobacillus plantarum 1-S27202 from tempe, Lactobacillus rhamnosus R23 and Pediococcus pentosaceus 1-A38 from human breast milk and a commercially available human isolates Lactobacillus acidophilus FNCC0051 in lowering cholesterol by in vitro and metabolizing the prebiotic oligosaccharide compounds. The effects of oligosaccharide compounds on the performance of the LAB isolates in lowering cholesterol were also evaluated. The tests were done in MRS based medium in vitro with or without oligosaccharides i.e. galactooligosacharrides (GOS), fructooligosaccharides (FOS), inulin, hydrolyzed inulin or combination of oligosaccharides as prebiotics. The results revealed that all isolates were able to reduce cholesterol in the medium, and the highest cholesterol reduction was observed for L. acidophilus FNCC0051 and L. rhamnosus R23. There are two different mechanism in the loweringof cholesterol; cholesterol assimilation and cholesterol binding on the cell surface. For the case of P. pentosaceus 1-A38, it involves the assimilation, while the other four isolates may involve cholesterol binding on the cell surface. In addition, the tested LAB's has different ability to use prebiotics, as shown by the reduction of total sugar in the medium. Oligosaccharides metabolism by L. acidophilus FNCC0051 and L. rhamnosus R23 resulted in several organic acid and SCFA with lactic acid produced as the largest proportion followed by acetic acid. Furthermore, the proportion of propionic and butyric acids were influenced by the type of isolates and carbon source. L. acidophilus FNCC 0051 was able to reduce cholesterol in the MRS based medium with oligosaccharides and their combination as carbon source and cholesterol reducing ability seems to involve both assimilation and cholesterol binding on the cell surface.