One major challenge in the field of tissue engineering was the creation of volumetric tissues and organs in vitro. To achieve this goal, the development of a three-dimensional vascular-like network that extended throughout the tissue engineered construct was essential to supply sufficient oxygen and nutrients to all of the cells in the constructs. For sufficient oxygenation and nutrition of the tissue-engineered constructs, the distance between each microvessel-like channel in the network should ideally be within 100-200 mu m. In addition, the medium or blood should be perfused through the microchannels as soon as possible after the seeding of cells into the templates (scaffolds) of the constructs. In the present study, we proposed a novel technique for fabricating an engineered vascular-like network that satisfied these two requirements. The network comprised assembled hollow alginate hydrogel microfibers with mammalian cells enclosed in the gel portions. We controlled the distance between each flow microchannel (hollow core portions and interspace of the microfibers) to be within 150 gm by using microfibers with a gel thickness of approximately 50 gm. Furthermore, we confirmed that medium could be perfused into the flow channels quickly (within 10 min) after immobilization of the cells in the assembly. A human hepatoblastoma cell line (HepG2) proliferated in the gel portions of the microfibers and maintained their specific function during perfusion culture for 7 days. These results showed that the novel vascular-like networks fabricated here had the potential to allow the creation of volumetric tissues in vitro. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.