PLGA (poly(lactic-co-glycolic acid)) is one of the most used biodegradable and biocompatible materials. Nanostructured PLGA even has great application potentials in tissue engineering. In this research, a fabrication technique for nanostructured PLGA membrane was investigated and developed. In this novel fabrication approach, an anodic aluminum oxide (AAO) film was use as the template; the PLGA solution was then cast on it; the vacuum air-extraction process was applied to transfer the nano porous pattern from the AAO membrane to the PLGA membrane and form nanostures on it. The cell culture experiments of the bovine endothelial cells demonstrated that the nanostructured PLGA membrane can double the cell growing rate. Compared to the conventional chemical-etching process, the physical fabrication method proposed in this research not only is simpler but also does not alter the characteristics of the PLGA. The nanostructure of the PLGA membrane can be well controlled by the AAO temperate.