Isolation of skeletal precursor cells

• US 7,863,045 B2
• Filed: 07/18/2008
• Issued: 01/04/2011
• Est. Priority Date: 10/06/1999
• Status: Expired due to Fees

• Current Assignee: Tigenix NV (Takeda Pharmaceutical Company Limited)
• Original Assignee: Tigenix NV (Takeda Pharmaceutical Company Limited)
• Inventors: Luyten, Frank, Dell'Accio, Francesco, De Bari, Cosimo
• Primary Examiner(s): Ton; Thaian N
• Application Number:
  US12/176,256
• Publication Number:
  US 20090123927A1
• Time in Patent Office: 900 Days
• Field of Search: 435/378, 435/7.21, 435/366, 435/372
• US Class Current: 435/378
• 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 identifying viable, expanded or passaged, committed skeletal precursor cells capable of differentiating into skeletal tissues comprising the step of:

detecting the expression of cartilage-derived morphogenic protein-1 (CDMP-1) in an in vitro cell culture of cells obtained from human periosteum, synovial membrane or bone marrow,

wherein the expression of CDMP-1 identifies skeletal precursor cells.

2. The method according to claim 1, wherein the expression of CDMP-1 is detected at the DNA, mRNA, cDNA or protein level and/or via the activity of a CDMP-1 promoter that is operably linked to a heterologous reporter gene.

3. The method according to claim 1, wherein the step of detecting CDMP-1 expression includes applying a binding agent for CDMP-1 to the in vitro cell culture.

4. 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.

5. A method of obtaining a homogenous culture of viable, expanded or passaged, committed skeletal precursor cells capable of differentiating into skeletal tissues comprising the steps of:

a) detecting the expression of cartilage-derived morphogenic protein-1 (CDMP-1) in an in vitro cell culture of cells obtained from human periosteum, synovial membrane or bone marrow; and

b) selecting or enriching CDMP-1 positive cells, thereby producing a homogenous culture of skeletal precursor cells capable of differentiating into skeletal tissue.

6. The method according to claim 5 wherein the expression of CDMP-1 is detected at the DNA, mRNA, cDNA or protein level and/or via the activity of a CDMP-1 promoter that is operably linked to a heterologous reporter gene.

7. The method according to claim 5, wherein the step of detecting CDMP-1 expression includes applying a binding agent for CDMP-1 to the in vitro cell culture.

8. The method according to claim 5, 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.