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Background: The extensive use of poliovaccines has eliminated the wild-type poliovirus in most parts of the world. These conditions were caused due to the utilization of oral polio vaccine (OPV) and inactive polio vaccine (IPV). Since most of the quality control tests for these vaccines are performed on cell beds sensitive to poliovirus, the identification of the most sensitive cell line to poliovirus is a necessity. Materials and Methods: Five monolayer cell lines (Vero, HeLa, Hep-2, MRC-5 and L20- B) were prepared in cell culture flasks (25 cm2). Then serial dilutions of three types of poliovirus with specified titers were added to each cell beds. The inoculated cells were then incubated at 33°C for 14 days and were monitored daily for the presence of cytopathic effects for polioviruses. Results: The results showed that the sensitivity of L20B cell line to polioviruses was more than the other cells. The result also indicated that the sensitivity of cells to poliovirus was declined in Hep-2, HeLa, MRC-5 and Vero cell lines, respectively. Conclusion: It can be concluded that the L20B, Hep-2 and HeLa cell lines, due to their higher sensitivity to triple poliovirus serotypes are considered for vaccine quality control tests. [ABSTRACT FROM AUTHOR]