We made micropatterned vascular endothelial cells, which have a regular capillary tube-like structure, on a bioactive hydrogel matrix. We applied a stamping method to transfer micropatterned bovine aortic endothelial cells to a growth factor-reduced basement membrane matrix (Matrigel) and type I collagen gel. In this study, we addressed the issues of how to accelerate cell transfer and the effective factors in doing so. We focused on the effects of the cell-substratum and cell-cell adhesiveness prior to applying cultured endothelial cells to a hydrogel matrix on cellular behavior under transfer printing. We found that individual cells cultured sparsely on substrata with different cell adhesivity transferred to Matrigel up to 40%, whereas cells cultured on patterned substrata having lines of 60 mu m in width, which involved cell-cell contacts, could transfer homogeneously to Matrigel within a few hours. The morphology of such cells changed from a tape-like monolayer into a thinner, tube-like structure. The speed and the ratio of transfer of micropatterned cells to Matrigel were affected by the period of cell culture on micropatterned substrata. We also found that the intensity of vascular endothelial cadherin staining at cell-cell junctions of micropatterned cells was correlated with cellular behavior when applying them to Matrigel, on which cells formed a tube-like structure or to which cells migrated individually. Furthermore, micropatterned cells made regular tube-like structures when applied to type I collagen gel. Such tube-like endothelial cells had good viability. These findings may be useful for creating in vitro angiogenesis assays and tissue-like constructs that include capillary-like networks of vascular endothelial cells.