Heart muscle-like cell sheet, three-dimensional construct, heart muscle-like tissue and process for producing the same

• US 20040009566A1
• Filed: 07/15/2003
• Published: 01/15/2004
• Est. Priority Date: 07/21/2000
• Status: Abandoned Application

• Current Assignee: CellSeed Inc.
• Original Assignee: CellSeed Inc.
• Inventors: Shimizu, Tatsuya, Okano, Teruo, Kikuchi, Akihiko, Yamato, Masayuki
• Application Number:
  US10/333,473
• Publication Number:
  US 20040009566A1
• Time in Patent Office: Days
• US Class Current: 435/174
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61L 27/3826 Muscle cells, e.g. smooth muscle cells
• CPC Class Codes: A61L 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: C12N 2539/10 Coating allowing for selective detachment of cells, e.g. thermoreactive coating
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells

Abstract

Cells are cultured on a cell culture support having a substrate surface coated with a temperature-responsive polymer whose upper or lower critical solution temperature in water is 0˜80° C., and subsequently, (1) the temperature of the culture solution is brought to above the upper critical solution temperature or below the lower critical solution temperature, and optionally, (2) the cultured cell sheet is brought into close contact with a polymer membrane and (3) the cell sheet is peeled off together with the polymer membrane. In this way, the myocardial cell sheet can be cultured three-dimensionally to construct a myocardium-like tissue.

Claims

1. A myocardium-like cultured cell sheet made of myocardial tissue cells that retain contracting and relaxing functions, intercellular electrical coupling and orientation.

3. The myocardium-like cultured cell sheet according to claim 1 or the three-dimensional structure according to claim 2, which have been peeled off from a substrate without being treated with a proteolytic enzyme.

4. The myocardium-like cultured cell sheet or three-dimensional structure according to claim 1, 2 or 3, which have been oriented in a specified direction by stretching them in the specified direction after they were peeled off from the substrate.

11. A myocardium-like tissue produced by a method in which the myocardium-like cell sheet or three-dimensional structure of myocardium-like cells according to any one of claims 1˜

4 and 10 is buried in the living body so that tubular cavities are formed of vascular endothelial cells endogenous in said myocardium-like cell sheet or three-dimensional structure of myocardium-like cells and/or the vascular endothelial cells in the tissue around the buried graft are allowed to grow inward and form tubular cavities, thereby forming blood vessels.

12. The myocardium-like cell sheet or three-dimensional structure of myocardium-like cells according to any one of claims 1˜

4 and 10 or the myocardium-like tissue according to claim 11, which are suitable for use in the treatment of heart disease and other circulatory organ related diseases or digestive organ related diseases.

13. A method of treating heart disease and other circulatory organ related diseases or digestive organ related diseases using the myocardium-like cell sheet or three-dimensional structure of myocardium-like cells according to any one of claims 1˜

4 and 10 or the myocardium-like tissue according to claim 11.

2. A three-dimensional structure of myocardium-like cultured cells that retain contracting and relaxing functions, as well as three-dimensional intercellular electrical coupling and orientation, said structure forming tubular cavities of vascular endothelial cells and/or having a single epicardium-like outer cell layer.

5. A process for producing a myocardium-like cell sheet, which comprises culturing cells on a cell culture support having a substrate surface coated with a temperature-responsive polymer whose upper or lower critical solution temperature in water is 0˜

80°

C., and subsequently;

(1) bringing the temperature of the culture solution to above the upper critical solution temperature or below the lower critical solution temperature, and optionally;

(2) bringing the cultured cell sheet into close contact with a polymer membrane; and

(3) peeling the cell sheet off together with the polymer membrane.

6. A process for producing a three-dimensional structure of myocardium-like cells, in which the myocardium-like cell sheet as obtained in claim 5 is again allowed to adhere to a cell culture support, a cell culture support coated with a temperature-responsive polymer, a polymer membrane or a cellular sheet and a plurality of the assemblies are piled up.

7. The process for producing a myocardium-like cell sheet according to claim 5 or the process for producing a three-dimensional structure of myocardium-like cells according to claim 6, wherein the step of peeling off does not involve treatment with a proteolytic enzyme.

8. The process for producing a myocardium-like cell sheet according to claim 5 or the process for producing a three-dimensional structure of myocardium-like cells according to claim 6, wherein the temperature-responsive polymer is poly(N-isopropylacrylamide).

9. The process for producing a myocardium-like cell sheet according to claim 5 or the process for producing a three-dimensional structure of myocardium-like cells according to claim 6, wherein the polymer membrane is selected from among a hydrophilized polyvinylidene difluoride membrane, polyurethane, a Spandex mesh and a stockinet-like material.

10. A myocardium-like cell sheet or a three-dimensional structure of myocardium-like cells that are produced by the process according to any one of claims 5˜

9.