This article describes a novel laser-directed microfabrication method carried out in aqueous solution for the organization of cell networks on a platform. A femtosecond (fs) laser was applied to a platform culturing PC12, HeLa, or normal human astrocyte (NHA) cells to manipulate them and to facilitate mutual connections. By applying an fs-laser-induced impulsive force, cells were detached from their original location on the plate, and translocated onto microfabricated cell-adhesive domains that were surrounded with a cell-repellent perfluoroalkyl (Rf) polymer. Then the fs-laser pulse-train was applied to the Rf polymer surface to modify the cell-repellent surface, and to make cell-adhesive channels of several μm in width between each cell-adhesive domain. PC12 cells elongated along the channels and made contact with others cells. HeLa and NHA cells also migrated along the channels and connected to the other cells. Surface analysis by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) confirmed that the Rf polymer was partially decomposed. The method presented here could contribute not only to the study of developing networks of neuronal, glial, and capillary cells, but also to the quantitative analysis of nerve function. To make channels for cell-to-cell connections on a platform, an intense femtosecond laser pulse train was focused on the perfluoroalkyl (Rf) polymer layer during the cell culture process. This treatment modified the Rf polymer surface to create a cell-adhesive region. Cells elongated or migrated along the modified area and made cell-cell connections.