Nanofiber scaffolds are widely used as the platform for three-dimensional culture of hepatocytes in vitro. The pore size of scaffolds plays an important role in promoting the infiltration and proliferation of hepatocyte. We show that the average pore size of electrospun scaffold increases from similar to 7.6 to 13.2 mu m, while the average fiber diameter decreases from similar to 2.0 to 1.5 mu m when collected by probe array collectors. Though increase in pore size decreases the tensile stress of scaffolds, it leads to enhance the proliferation and attachment of hepatocytes. Specifically, a 6 x 6 array scaffold which was prepared by probe collector was orderly arrayed. Compared with the conventional scaffold, the pore size of the arrayed scaffold doubled and the hydrophilicity was improved. When HepG(2) cells were seeded on the arrayed scaffold, cells showed superior adhesion ability, better cell morphology and three-dimensional growth. These results indicated that the ordered 6 x 6 array scaffold has the potential as a suitable substratum for in vitro culture of hepatocytes.