Fabrication of 3D vascularized bone graft is a major challenge in bone tissue engineering.  Here, we propose a fabrication strategy of autologous bone-like structures using bone beads in vitro.

　Bone beads were fabricated by micro-dispensing the collagen gel solution embedded mesenchymal stem cells (MSCs) onto a hydrophobic surface.  After 24 h of culture, the beads self-formed cell-dense spherical tissues by cell traction force.  During 14 d of osteogenic differentiation culture, MSCs embedded in the beads were differentiated to osteoblast and produced calcium phosphate.  Human umbilical vein endothelial cells (HUVECs) adhering on the calcified bone beads and collagen gel were poured into micro-chamber.  During 2d of culture, encapsulated HUVECs formed microvascular around the bone beads.  In this way, we fabricated autologous bone-like structures which have microvascular, osteoblast, and bone substrate (collagen and calcium phosphate).