Scaffold-free self-organized 3D synthetic tissue

• US 9,370,606 B2
• Filed: 02/16/2007
• Issued: 06/21/2016
• Est. Priority Date: 08/01/2003
• Status: Active Grant

• Current Assignee: Two Cells Co. Ltd.
• Original Assignee: Two Cells Co. Ltd.
• Inventors: Nakamura, Norimasa, Yoshikawa, Hideki, Ando, Wataru
• Primary Examiner(s): DAVIS, RUTH A
• Application Number:
  US11/676,172
• Publication Number:
  US 20080004713A1
• Time in Patent Office: 3,413 Days
• Field of Search: 424/400, 424/422, 424/423, 424/93.1, 435/325
• US Class Current: 1/1
• 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 27/3633 Extracellular matrix [ECM]
• CPC Class Codes: A61L 27/3843 Connective tissue
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage

Abstract

The present invention can be used for actual implantation surgery without a scaffold. The present invention provides a synthetic tissue or complex which can be produced by culture and has a high level of differentiation ability. The present invention also provides a therapy and medicament for repairing and/or regenerating tissue using replacement and covering. By culturing cells under specific culture conditions such that medium contains an extracellular matrix synthesis promoting agent, the cells are organized and are easily detached from a culture dish. The present invention was achieved by finding such a phenomenon. In addition, the self contraction of the tissue can be regulated by culturing the tissue in a suspended manner. Therefore, it is possible to regulate the three-dimensional shape of the tissue. The present invention also provides a method for producing an implantable synthetic tissue which does not require a plurality of monolayer cell sheets assembled to form a three-dimensionally structured synthetic tissue. The present invention is characterized by richness in adhesion molecules, nonnecessity of additional fixation at an implantation site, and good biological integration.

Claims

1. An implantable synthetic autograft or allograft tissue for implantation in a recipient, which is substantially made of (i) cells that are autologous or allogeneic relative to the recipient and which thereby constitute the synthetic autograft or allograft tissue, said cells being selected from the group consisting of myoblasts, mesenchymal stem cells, adipocytes, synovial cells and bone marrow cells;

and (ii) an extracellular matrix (ECM) derived from the cells constituting the synthetic autograft or allograft tissue, said extracellular matrix comprising fibronectin, collagen I, collagen III, and vitronectin,

wherein the synthetic autograft or allograft tissue is free of scaffolds and comprises multiple layers of said cells, and is formed by a method that comprises detaching a cultured monolayer of said cells from a culture container without contacting the cells with a protein degrading enzyme, under conditions and for a time sufficient to obtain a detached, self-contracted synthetic tissue,

wherein the fibronectin, collagen I, collagen III, and vitronectin are evenly distributed in the synthetic autograft or allograft tissue,

wherein the fibronectin, collagen I, collagen III, and vitronectin three-dimensionally wrap the cells, and

wherein the implantable synthetic autograft or allograft tissue has integration ability with its surroundings when implanted and has sufficient strength to provide self-supporting ability.

2. The implantable synthetic autograft or allograft tissue of claim 1, wherein the extracellular matrix is integrated throughout the tissue.

3. The implantable synthetic autograft or allograft tissue of claim 1 which comprises 10 or more layers of said cells.

4. The implantable synthetic autograft or allograft tissue of claim 1 which has a thickness of at least about 50 μ

m and is dimensioned so as to fit an injured site in a tissue or organ.