In this study, we have prepared a rubbed fluorinated polyimide membrane using a rubbing machine with a rubbing cloth, followed by ion‐irradiation, and formed hepatocyte spheroids on the resulting patterned membrane. The re‐seeding of the spheroids, that is, spheroid building, enhanced the biochemical functions of the spheroids by optimization of the culture conditions. Furthermore, the tubular formation of bovine aortic endothelial cells (BAECs) on the resulting patterned membrane has been demonstrated. The pre‐network structure of BAECs was observed, forming the network and tube formation after gel culture. The combination of the built spheroids and the resulting BAEC network is expected to be a unique method to regenerative damaged tissues. Copyright © 2011 John Wiley & Sons, Ltd.