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The long-term goals of this project are to isolate and characterize
stem cells from human skeletal muscle, and test their ability
to correct muscular deficiencies in-vivo. These studies will
require the use of the fluorescence-activated cell sorter
and microarray techniques to purify human muscle cells and
identify the genes they specifically express. Human muscle
stem cells will be introduced into the circulation of different
animal models of muscle disorders and their ability to target
and repair damaged muscle will be assessed.
AIMS:
- To optimize the isolation of muscle stem cells (SP cells)
in humans.
- To use chip-based mRNA expression arrays to identify genes
expressed by human muscle SP cells.
- To analyze candidate genes from gene chip technology using
other conventional techniques.
- To optimize conditions to propagate and culture human
SP cells in vitro.
- To evaluate the differentiation potential of human muscle
SP cells in vivo.
These studies are essential to further our basic knowledge
on the existence of muscle stem cells in humans. The identification
of candidate genes that are uniquely expressed by human muscle
stem cells will help in understanding how muscle stem cells
differ from more committed myoblasts, and start to unravel
why muscle stem cells (at least from previous mouse studies)
can differentiate into bone marrow. Further, exploring methods
to propagate muscle stem cells will be crucial to obtaining
large numbers of cells for characterization experiments as
well as in vivo studies. These in vivo studies are aimed to
test whether human muscle stem cells can be safely used as
vehicles to systemically deliver genes to skeletal muscle.
The hope is to be able to extend the practical use of muscle
stem cells in the development of a therapy for human muscle
disorders.
Photographs from our lab:
| Figure 1. Dystrophin-positive
myofiber (green) detected in an mdx (muscular dystrophy)
mouse injected with male mouse wild-type SP (side population)
cells. Nuclei are stained in blue by DAPI. The donor nucleus
within the dystrophin-positive myofiber has a red-hybridization
signal (Y-chromosome). |
Figure 1. |
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