Method for preparing engineered tissue
View Patent ↗A method for preparing a human or animal tissue by applying a compressive force to a stack of sheets of living tissue thereby inducing adjacent layers to fuse or adhere to each other. The force is applied in direction normal to the surface of the tissue. A multi-layer tissue produced by the method described above can also possess at least two different types of sheets and/or consist essentially of between two and twelve sheets of living tissue. The method can also be used to prepare a planar tissue that can further be incorporated in a multi-layer tissue construct. The methods and tissues described herein are useful for the preparation of engineered tissues.
1. A method for preparing a human or animal tissue from at least one sheet of living tissue, the method comprising the steps of: (a) arranging said at least one sheet of living tissue to form a multi-layer stack of living tissue; and (b) applying a compressive force in a direction normal to the surface of the multi-layer stack of living tissue with a force-applying means at a pressure and for an amount of time sufficient to compress layers of tissue together for inducing adjacent layers of tissue to fuse or adhere to each other, wherein said force-applying means in step (b) comprises an adjustable weighted device suitable for applying substantially evenly-distributed pressure to said multi-layer stack of living tissue, said weighted device being at least partially permeable to tissue-culture medium.
2. The method of claim 1 , wherein said multi-layer stack is arranged on a substantially flat support.
3. The method of claim 1 , wherein said multi-layer stack of living tissue in step (a) is formed by superimposing two or more sheets of living tissue.
4. The method of claim 1 , wherein said multi-layer stack of living tissue is formed by folding a sheet of living tissue upon itself.
5. The method of claim 1 , further comprising a step of anchoring said multi-layer stack of living tissue with anchoring means before said step (b) of applying a force, wherein said anchoring means applies sufficient tension across said multi-layer stack of living tissue to prevent shrinkage and/or maintain cellular differentiation and/or induce fiber orientation.
6. The method of claim 5 , wherein said anchoring means comprises a multiplicity of spaced apart weights or ingots arranged substantially around the perimeter of said multi-layer stack of living tissue.
7. The method of claim 1 , wherein the multi-layered stack of living tissue in step (a) is formed by rolling a sheet of living tissue on a tubular support.
8. The method of claim 7 , wherein said force-applying means in step (b) comprises a tissue-culture medium permeable elastic sleeve.
9. The method of claim 1 , wherein cells are obtained from said at least one sheet of living tissue, said at least one sheet of living tissue is obtained via biopsy.
10. The method of claim 1 , wherein said at least one sheet of living tissue is obtained by culturing cells in vitro.
11. The method of claim 10 , wherein said cells are selected from the group consisting of embryonic stem cells, post-natal stem cells, adult stem cells, mesenchymal cells, hepatocytes, islet cells, parenchymal cells, osteoblasts and other cells forming bone or cartilage, and nerve cells.
12. The method of claim 11 , wherein said mesenchymal cells are selected from the group consisting of fibroblasts, interstitial cells, endothelial cells, smooth muscle cells, skeletal muscle cells, myocytes, chrondocytes, adipocytes, fibromyoblasts, and ectodermal cells.
13. The method of claim 1 , wherein said at least one sheet of living tissue is selected from the group consisting of a skin tissue, a corneal tissue, a cardiac valve tissue, a connective tissue and a mesenchymal tissue.
14. A multi-layer tissue made according to the method of claim 1 , wherein said multi-layer tissue comprises at least two different types of sheets of living tissue.
15. A multi-layer tissue according to claim 14 , consisting essentially of between two sheets and twelve sheets of living tissue.
16. A multi-layer tissue according to claim 14 , consisting essentially of between three sheets and nine sheets of living tissue.
17. A multi-layer tissue according to claim 14 , wherein said tissue has a thickness of about 0.01 mm to about 0.5 mm.
18. A multi-layer tissue according to claim 17 , wherein said tissue has a thickness of about 0.03 mm to about 0.45 mm.