ISOLATION OF PRECURSOR CELLS AND THEIR USE FOR TISSUE REPAIR

• US 20090123927A1
• Filed: 07/18/2008
• Published: 05/14/2009
• Est. Priority Date: 10/06/1999
• Status: Active Grant

• Current Assignee: Tigenix NV (Takeda Pharmaceutical Company Limited)
• Original Assignee: Tigenix NV (Takeda Pharmaceutical Company Limited)
• Inventors: DELL'ACCIO, Francesco, LUYTEN, Frank, DE BARI, Cosimo
• Granted Patent: US 7,863,045 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3817 Cartilage-forming cells, e.g. pre-chondrocytes
• CPC Class Codes: A61L 27/3843 Connective tissue
• CPC Class Codes: A61L 27/3886 comprising two or more cell types
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2502/1317 Chondrocytes
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

Cartilage-derived morphogenetic protein CDMP-1 or a transforming growth factor β having at least 80% homology with CDMP-1, or a factor co-expressed and/or co-detectable therewith, is used as a marker of skeletal precursor cells from any part of a mammalian body.

Claims

1. A method of positively identifying viable, expanded or passaged, committed, pluripotent skeletal precursor cells that have entered a post-natal differentiation pathway leading to skeletal or connective tissues comprising the steps of isolating mammalian cells into a cell culture in vitro, and detecting the expression by said cells of a positive embryonic marker which is cartilage-derived morphogenic protein CDMP-1.

2. The method according to claim 1 which comprises detecting the positive marker at the DNA, mRNA, cDNA or the protein level and/or via the activity of a promoter directing/regulating this gene expression, operably linked to heterologous reporter gene.

3. The method according to claim 1 wherein said precursor cells are isolated from a joint interzone of a mammal.

4. The method according to claim 1, wherein the step of detecting the presence of the positive marker includes applying a binding agent for the positive marker to an isolated source of cell having the precursor cells, the marker positively identifying the cell.

5. The method according to claim 1, wherein said step of isolating mammalian cells comprises isolating cells from periosteum, bone marrow, or synovial membrane.

6. The method according to claim 1, which further comprises detecting the absence of expression of a marker selected from the group consisting of FGFR3, type II collagen, type IX collagen, type X collagen, type XI collagen, and BMP-2

7. A method for obtaining a homogenous culture of viable, expanded or passaged, committed, pluripotent skeletal precursor cells that have entered a post-natal differentiation pathway leading to skeletal or connective tissues comprising the steps of isolating mammalian cells into a cell culture in vitro, and determining the expression by said culture of a positive embryonic marker which is cartilage-derived morphogenic protein CDMP-1.