Patent ID: 8076136
Filing Date: 2011-12-13
Classification: A61L,C12N

Abstract:
1. A method for promoting formation of a three-dimensional osteogenic cell aggregate ex vivo, the method comprising: (a) introducing osteoclast precursors and osteoblasts into a cylindrical culture vessel that rotates about a central horizontal axis, wherein said cylindrical culture vessel contains a matrix-free culture medium, wherein the matrix-free culture medium does not comprise an exogenous scaffolding material that guides formation of the three-dimensional osteogenic cell aggregate, (b) co-culturing said osteoblasts and said osteoclast precursors in said cylindrical culture vessel during horizontal rotation at a rate of between 1 and 2 rotations per minute to promote scaffold-free interaction, aggregation and organization of said osteoclast precursors and said osteoblasts until the three dimensional tissue constructs are formed, (c) further culturing, the tome dimensional aggregate of step (b) in a matrix-free mineralization culture medium in said cylindrical culture vessel at a higher average rate of horizontal rotation than the average rate of horizontal rotation during step (b) to increase perfusion and thereby to form mature three dimensional mineralized osteogenic cell aggregate composed of osteoblasts and osteoclasts differentiated from osteoclast precursors, wherein mineralized three dimensional osteogenic cell aggregate comprise an outer zone comprising osteoclasts surrounding an inner porous core comprising osteoblasts and/or osteocytes encased in a mineralized extracellular matrix.