Process for ex vivo formation of mammalian bone and uses thereof
AssigneeThe Regents of the University of Michigan
InventorsSujata Kale, Michael W. Long
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Abstract
The present invention concerns methods for the ex vivo formation of mammalian bone and subsequent uses of the bone. A critical and distinguishing feature of the present invention are defined tissue culture conditions and factors resulting in the formation of bone cell spheroids. The invention also provides for methods of implanting into subjects the ex vivo formed bone. Also described are methods for genetically altering the bone cell spheroids to affect bone formation, identification of candidate modulators of bone formation, and identification of genes involved in bone formation.
Claims31
A method for producing bone ex vivo, comprising the steps of:
- a) obtaining (i) osteoblast cells, (ii) preosteoblast cells or (iii) bone precursor cells that express low amounts of bone proteins and exhibit a low degree of internal complexity;
- b) culturing said cells under serum free conditions in the presence of one or more osteogenic growth factors and at cell densities which produce a bone cell spheroid of approximately 3000 to 100,000 cells;
- and
- c) maintaining said bone cell spheroid under conditions which produce bone within said bone cell spheroid.
A method of providing bone tissue to a mammal, comprising obtaining a bone cell spheroid of approximately 3000 to 100,000 bone cells and implanting the bone cell spheroid into said mammal.
A method for producing and using bone comprising the steps of:
- a) obtaining (i) osteoblast cells, (ii) preosteoblast cells or (iii) bone precursor cells that express low amounts of bone proteins and exhibit a low degree of internal complexity;
- b) culturing said cells under serum free conditions in the presence of one or more osteogenic growth factors and at cell densities which produce a bone cell spheroid of approximately 3000 to 100,000 cells;
- c) maintaining the bone cell spheroid under conditions which produce bone;
- and,
- d) removing the cellular elements from the formed bone and using resulting bone in vivo.
A bone cell spheroid made by the process of:
- a) obtaining (i) osteoblast cells, (ii) preosteoblast cells or (iii) bone precursor cells that express low amounts of bone proteins and exhibit a low degree of internal complexity;
- and
- b) culturing said cells under serum free conditions in the presence of one or more osteogenic growth factors and at cell densities which produce a bone cell spheroid of approximately 3000 to 100,000 cells.
Dependencies are inferred from the claim text.