A microcarrier is used for the three-dimensional (3D) culture of adhesion-dependent mammalian cells. We developed a novel microcarrier by binding ProNectin F, an artificial cell adhesive protein synthesized by genetically engineered Escherichia coil to a polyacrylic superabsorbent polymer. The microcarrier is characterized by containing no animal-derived components. The serum-free culture of Vero cells for vaccine production using the microcarrier increased the number of Vero cells by approximately 30% compared with the existing dextran beads coated with porcine Type I collagen, which resulted in approximately a 30% to 40% increase in the infectivity titer of the Sabin 2 strain of poliovirus. These results suggested that the developed microcarrier should be unprecedented in permitting high-yield vaccine production by means of a serum-free culture. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.