Monoclonal antibody capable of binding integrin alpha 10 beta 1
The present invention provides a monoclonal antibody or a fragment thereof binding to the extracellular I-domain of integrin alpha10beta1 and a hybridoma cell line deposited at the Deutsche Sammlung von Microorganismen and Zellkulturen GmbH under the accession number DSM ACC2583. Furthermore, the present invention also provides a monoclonal antibody or a fragment thereof binding to the extracellular I-domain of integrin alpha10beta1 produced by the hybridoma cell line deposited. Methods and uses of said antibody or a fragment thereof in identifying and selecting cells of a chondrogenic nature for treatment purposes, in particular for the identification and isolation of chondrocytes, mesenchymal progenitor cells and embryonic stem cells for tissue engineering of cartilage, or for identifying diagnostic and therapeutic tools in studying the biological role and the structural/functional relationships of the integrin alpha10beta1 with its various extracellular matrix ligands are also included.
1. A method for isolating a population of mammalian mesenchymal stem cells, the method comprising the steps of:
a) providing a cell suspension comprising mammalian mesenchymal stem cells,
b) contacting the cell suspension with a monoclonal antibody or a fragment thereof capable of binding specifically to the extracellular I-domain of the integrin alpha10 chain, under conditions wherein said monoclonal antibody or fragment thereof forms an antibody-antigen complex with the extracellular domain of integrin alpha10beta1, wherein said monoclonal antibody is antibody 365 produced by a hybridoma cell line deposited at the Deutsche Sammlunq von Microoraanismen und Zellkulturen GmbH under the accession number DSM ACC2583, and
c) separating cells binding to the monoclonal antibody or fragment thereof in b), wherein separation is achieved mechanically via a solid support bound directly or indirectly to the monoclonal antibody or fragment thereof, or by cell sorting via a label bound directly or indirectly to the monoclonal antibody or fragment thereof,
thereby producing a population of mammalian mesenchymal stem cells.
2. The method according to claim 1 , wherein the monoclonal antibody or fragment thereof is linked to a solid phase.
3. The method according to claim 1 , wherein the solid phase is beads.
4. The method according to claim 1 , wherein the mammalian cells are human cells.
5. The method according to claim 1 , wherein the mammalian cells are murine cells.
6. The method of claim 1 , wherein separation is achieved mechanically via beads bound directly or indirectly to the monoclonal antibody or fragment thereof.
7. The method of claim 1 , wherein separation is achieved by fluorescence activated cell sorting (FACS).
8. The method of claim 1 , further comprising recovering cells binding to the monoclonal antibody or fragment thereof in c) from said antibody or fragment thereof.
9. The method of claim 1 , wherein the isolated population of mammalian mesenchymal stem cells is free from the monoclonal antibody or fragment thereof.
10. A method for detecting a mesenchymal stem cell in a sample, the method comprising the steps of:
a) providing a sample cell suspension comprising a mesenchymal stem cell,
b) contacting said sample cell suspension with a monoclonal antibody or a fragment thereof capable of binding specifically to the extracellular I-domain of the integrin alpha10 chain, wherein said monoclonal antibody is antibody 365 produced by a hybridoma cell line deposited at the Deutsche Sammlung von Microorganismen and Zellkulturen GmbH under the accession number DSM ACC2583,
c) incubating the sample cell suspension and the monoclonal antibody or fragment thereof under conditions wherein said monoclonal antibody or fragment thereof forms an antibody-antigen complex with the extracellular domain of integrin alpha10beta1 on a mesenchymal stem cell, and
d) detecting the monoclonal antibody or fragment thereof of c) via a detectable moiety associated directly or indirectly with said monoclonal antibody or fragment thereof.
11. The method of claim 10 , wherein the monoclonal antibody or fragment thereof of c) is conjugated to a detectable moiety, wherein said detectable moiety is capable of producing, either directly or indirectly, a detectable signal, and wherein said detectable signal is used for detection of the monoclonal antibody or fragment thereof of c).
12. The method of claim 10 , wherein further a second antibody or an antigen-binding fragment thereof is added to the sample cell suspension, wherein said second antibody or fragment thereof is labeled and binds to the monoclonal antibody or fragment thereof of c), wherein said labeled second antibody or fragment thereof is used for detection of the monoclonal antibody or fragment thereof of c).