Molecular marker for muscle stem cells
AssigneeThe General Hospital Corporation, a Massachusetts corporation
InventorJeffrey B. Miller
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Abstract
In accordance with the invention, Bcl-2 expression is a molecular marker for muscle stem cells. Thus, the invention provides methods for identifying and isolating muscle stem cells. In addition, the invention provides methods for determining whether a test compound modulates muscle stem cell differentiation and/or proliferation. Finally, the invention provides methods for expressing an exogenous coding sequence in a muscle stem cell.
Claims20
A method for identifying a muscle stem cell, the method comprising providing a sample comprising a myogenic cell, and detecting activity of a Bcl-2 promoter within the myogenic cell as an indication that the myogenic cell is a muscle stem cell.
A method for determining whether a test compound modulates muscle stem cell differentiation, the method comprising:
- (a) providing a myogenic cell identified as a muscle stem cell;
- (b) contacting the muscle stem cell with the test compound;
- and
- (c) detecting a change in differentiation of the muscle stem cell as an indication-that the test compound modulates muscle stem cell differentiation.
A method for determining whether a test compound modulates muscle stem cell proliferation, the method comprising:
- (a) providing a myogenic cell identified as a muscle stem cell;
- (b) contacting the muscle stem cell with the test compound;
- and
- (c) detecting a change in proliferation of the muscle stem cell as an indication that the test compound modulates muscle stem cell proliferation.
A method for producing a population of cells enriched for muscle stem cells relative to a reference population of cells, the method comprising:
- providing a reference population of cells comprising a plurality of muscle stem cells and at least one cell other than a muscle stem cell;
- introducing into the reference population of cells a genetic construct comprising a Bcl-2 promoter operably linked to a gene encoding a marker protein that is heterologous to wild-type cells of the reference population, thereby producing a transfected population of cells;
- and
- selecting from the transfected population of cells those cells that express the marker protein, thereby producing a population of cells enriched for muscle stem cells.
A method for producing a population of living cells enriched for muscle stem cells relative to a reference population of cells, the method comprising:
- providing a reference population of living cells comprising a plurality of muscle stem cells that express Bcl-2 and at least one cell other than a muscle stem cell;
- and
- treating the reference population of cells to induce apoptosis in cells that do not express Bcl-2, thereby producing a population of living cells enriched for muscle stem cells.
A method for expressing an exogenous coding sequence in a muscle stem cell, the method comprising:
- (a) providing a myogenic cell identified as a muscle stem cell;
- (b) introducing into the muscle stem cell a genetic construct comprising an exogenous coding sequence operably linked to a muscle stem cell-active promoter, to produce a transfected muscle stem cell;
- and
- (c) maintaining the transfected muscle stem cell under conditions permitting expression of the exogenous coding sequence.
Dependencies are inferred from the claim text.