Abstract: Cell culture has played an important role in virology. It provides a platform for the detection and isolation of viruses as well as for the biochemistry and molecular biology study of viruses. In the present study, a new system that could support multiple different cell lines to be simultaneous cultured in one dish was developed. In the system, each cell line was cultured in an isolated zone in the same dish or well and the system is therefore called isolated co-cultured system. The usefulness of this novel approach for virus isolation was demonstrated using a model system based on adenovirus.