Microorganisms and their by-products are considered to be the major causes of pulpal and periapical pathosis. The role of microorganisms in endodotic infection has been studied regarding the prevalence of particular organisms found in root canal and periapical lesions. The aim of this study was to investigate the effect of culture supernatants of several oral microorganims isolated from infected root canals on the viability of cultured cell lines using colorimetirc assay. S. simulans, S. sciuri, E. faecium, S. intermedius, S. mitis, S. sanguis and S. uberis were incubated in Todd-Hewitt broth for 16 hours. 20 and 100ul of filtered bacterial cell culture supernatants were added to MK and Hep-2 cells. Cell viability was measured using MIT colorimetric assay. 20ul and 100u1 of S. sanguis supernatant showed significant cytotoxicity compared to control on MK cells. 100ul of S. sanguis supernatant significantly depressed viability of HEp-2 cells. E. faecium and S. intermedius did not affect the viability of MK and HEp-2 cells.