Background: Skeletal muscle contains populations of multipotent adult stem cells also referred to as muscle–derivedstem cells.Aim: The main goal of this study was to isolate and culture human adult stem cells from skeletal muscle and characterizethem.Methods: Muscle–derived stem cells were isolated from biopsy specimens of femoral muscle. The cells werecultured in Dulbecco's modifi ed Eagle's minimal essential medium supplemented with 10% fetal calf serum and gentamycin.When they reached confl uence, they were sub–passaged up to the third passage. Cells from the last passagewere prepared for TEM analysis. Production of ?–actin and desmin was confi rmed by histochemistry. Moreover, thephenotypic characterization was performed.Results: Primary isolated muscle–derived stem cells had a fi broblast–like shape. During subsequent passages theymaintained this morphology. TEM analysis showed typical ultrastructural morphology of mesenchymal stem cells.They had large pale nuclei with a large amount of euchromatine. Nuclei were irregular with noticeable nucleoli. Dilatedcisterns of rough endoplasmic reticulum were present in cytoplasm. In certain parts of the cytoplasm there were aggregatesof granules of glycogen. The products of cells were actively secreted into the extracellular matrix. They expressed?–actin and desmin. The results of phenotypic characterization showed that almost all analyzed cells were CD13,CD34, CD56 positive and CD45 negative. Moreover, they did not express anti–human fi broblast surface protein.Conclusions: Muscle–derived stem cells exhibited typical characteristics typical for mesenchymal stem cells. Afteranalysis of their diff erentiation potential they could be used in tissue engineering and regenerative medicine.

Institute of Medical Biology and Genetics Faculty of Medicine Comenius University Bratislava

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