Original Article

Clonal Isolation of Muscle-derived Cells Capable of Enhancing Muscle Regeneration and Bone Healing

Correspondence to: Johnny Huard, Director: Growth and Development Laboratory, Department of Orthopaedic Surgery and Molecular Genetics and Biochemistry, Children's Hospital of Pittsburgh and University of Pittsburgh, Pittsburgh, PA 15261.

Several recent studies suggest the isolation of stem cells in skeletal muscle, but the functional properties of these muscle-derived stem cells is still unclear. In the present study, we report the purification of muscle-derived stem cells from the mdx mouse, an animal model for Duchenne muscular dystrophy. We show that enrichment of desmin⁺ cells using the preplate technique from mouse primary muscle cell culture also enriches a cell population expressing CD34 and Bcl-2. The CD34⁺ cells and Bcl-2⁺ cells were found to reside within the basal lamina, where satellite cells are normally found. Clonal isolation and characterization from this CD34⁺Bcl-2⁺ enriched population yielded a putative muscle-derived stem cell, mc13, that is capable of differentiating into both myogenic and osteogenic lineage in vitro and in vivo. The mc13 cells are c-kit and CD45 negative and express: desmin, c-met and MNF, three markers expressed in early myogenic progenitors; Flk-1, a mouse homologue of KDR recently identified in humans as a key marker in hematopoietic cells with stem cell-like characteristics; and Sca-1, a marker for both skeletal muscle and hematopoietic stem cells. Intramuscular, and more importantly, intravenous injection of mc13 cells result in muscle regeneration and partial restoration of dystrophin in mdx mice. Transplantation of mc13 cells engineered to secrete osteogenic protein differentiate in osteogenic lineage and accelerate healing of a skull defect in SCID mice. Taken together, these results suggest the isolation of a population of muscle-derived stem cells capable of improving both muscle regeneration and bone healing.

Key Words: dystrophin, gene transfer, BMP-2, stem cells, bone formation

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• Shi, X., Garry, D. J. (2006). Muscle stem cells in development, regeneration, and disease.. Genes Dev. 20: 1692-1708 [Abstract] [Full Text]  
• Anderson, J. E. (2006). The satellite cell as a companion in skeletal muscle plasticity: currency, conveyance, clue, connector and colander. J. Exp. Biol. 209: 2276-2292 [Abstract] [Full Text]  
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• Franceschi, R.T. (2005). Biological Approaches to Bone Regeneration by Gene Therapy. J. Dent. Res. 84: 1093-1103 [Abstract] [Full Text]  
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• Hodgetts, S. I., Grounds, M. D. (2003). Irradiation of dystrophic host tissue prior to myoblast transfer therapy enhances initial (but not long-term) survival of donor myoblasts. J. Cell Sci. 116: 4131-4146 [Abstract] [Full Text]  
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• Munoz-Elias, G., Woodbury, D., Black, I. B. (2003). Marrow Stromal Cells, Mitosis, and Neuronal Differentiation: Stem Cell and Precursor Functions. Stem Cells 21: 437-448 [Abstract] [Full Text]  
• Howell, J. C., Yoder, M. C. (2003). Adult Stem Cell Plasticity Defined. NeoReviews 4: e181-186 [Full Text]  
• McLoon, L. K., Wirtschafter, J. (2003). Activated Satellite Cells in Extraocular Muscles of Normal Adult Monkeys and Humans. IOVS 44: 1927-1932 [Abstract] [Full Text]  
• Peterson, L., Minas, T., Brittberg, M., Lindahl, A. (2003). Treatment of Osteochondritis Dissecans of the Knee with Autologous Chondrocyte Transplantation: Results at Two to Ten Years. JBJS 85: 17-24 [Abstract] [Full Text]  
• Tallheden, T., Dennis, J. E., Lennon, D. P., Sjogren-Jansson, E., Caplan, A. I., Lindahl, A. (2003). Phenotypic Plasticity of Human Articular Chondrocytes. JBJS 85: 93-100 [Abstract] [Full Text]  
• Wada, M. R., Inagawa-Ogashiwa, M., Shimizu, S., Yasumoto, S., Hashimoto, N. (2003). Generation of different fates from multipotent muscle stem cells. Development 129: 2987-2995 [Abstract] [Full Text]  
• Steffel, J., Wernig, M., Knauf, U., Kumar, S., Wiestler, O. D., Wernig, A., Brustle, O. (2003). Migration and Differentiation of Myogenic Precursors Following Transplantation into the Developing Rat Brain. Stem Cells 21: 181-189 [Abstract] [Full Text]  
• Ito, C. Y., Li, C. Y. J., Bernstein, A., Dick, J. E., Stanford, W. L. (2003). Hematopoietic stem cell and progenitor defects in Sca-1/Ly-6A-null mice. Blood 101: 517-523 [Abstract] [Full Text]  
• Jankowski, R. J., Deasy, B. M., Cao, B., Gates, C., Huard, J. (2002). The role of CD34 expression and cellular fusion in the regeneration capacity of myogenic progenitor cells. J. Cell Sci. 115: 4361-4374 [Abstract] [Full Text]  
• Charge, S. B. P., Brack, A. S., Hughes, S. M. (2002). Aging-related satellite cell differentiation defect occurs prematurely after Ski-induced muscle hypertrophy. Am. J. Physiol. Cell Physiol. 283: C1228-C1241 [Abstract] [Full Text]  
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• Tamaki, T., Akatsuka, A., Yoshimura, S., Roy, R. R., Edgerton, V. R. (2002). New Fiber Formation in the Interstitial Spaces of Rat Skeletal Muscle During Postnatal Growth. J. Histochem. Cytochem. 50: 1097-1111 [Abstract] [Full Text]  
• Hannallah, D., Peterson, B., Lieberman, J. R., Fu, F. H., Huard, J. (2002). Gene Therapy in Orthopaedic Surgery. JBJS 84: 1046-1061 [Full Text]  
• Qu-Petersen, Z., Deasy, B., Jankowski, R., Ikezawa, M., Cummins, J., Pruchnic, R., Mytinger, J., Cao, B., Gates, C., Wernig, A., Huard, J. (2002). Identification of a novel population of muscle stem cells in mice: potential for muscle regeneration. J. Cell Biol. 157: 851-864 [Abstract] [Full Text]  
• Tamaki, T., Akatsuka, A., Ando, K., Nakamura, Y., Matsuzawa, H., Hotta, T., R Roy, R., Edgerton, V. R. (2002). Identification of myogenic-endothelial progenitor cells in the interstitial spaces of skeletal muscle. J. Cell Biol. 157: 571-577 [Abstract] [Full Text]  
• Minasi, M. G., Riminucci, M., De Angelis, L., Borello, U., Berarducci, B., Innocenzi, A., Caprioli, A., Sirabella, D., Baiocchi, M., De Maria, R., Boratto, R., Jaffredo, T., Broccoli, V., Bianco, P., Cossu, G. (2002). The meso-angioblast: a multipotent, self-renewing cell that originates from the dorsal aorta and differentiates into most mesodermal tissues. Development 129: 2773-2783 [Abstract] [Full Text]  
• Musgrave, D. S., Fu, F. H., Huard, J. (2002). Gene Therapy and Tissue Engineering in Orthopaedic Surgery. J Am Acad Orthop Surg 10: 6-15 [Abstract] [Full Text]  
• Deasy, B.M., Qu-Peterson, Z., Greenberger, J.S., Huard, J. (2002). Mechanisms of Muscle Stem Cell Expansion with Cytokines. Stem Cells 20: 50-60 [Abstract] [Full Text]  
• Mitchell, P. O., Pavlath, G. K. (2001). A muscle precursor cell-dependent pathway contributes to muscle growth after atrophy. Am. J. Physiol. Cell Physiol. 281: C1706-C1715 [Abstract] [Full Text]  
• Gerhart, J., Bast, B., Neely, C., Iem, S., Amegbe, P., Niewenhuis, R., Miklasz, S., Cheng, P. F., George-Weinstein, M. (2001). MyoD-positive myoblasts are present in mature fetal organs lacking skeletal muscle. J. Cell Biol. 155: 381-392 [Abstract] [Full Text]  
• Hawke, T. J., Garry, D. J. (2001). Myogenic satellite cells: physiology to molecular biology. J. Appl. Physiol. 91: 534-551 [Abstract] [Full Text]  
• Lee, J. Y., Musgrave, D., Pelinkovic, D., Fukushima, K., Cummins, J., Usas, A., Robbins, P., Fu, F. H., Huard, J. (2001). Effect of Bone Morphogenetic Protein-2-Expressing Muscle-Derived Cells on Healing of Critical-Sized Bone Defects in Mice. JBJS 83: 1032-1039 [Abstract] [Full Text]  

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