Danisovic Lubos, Varga Ivan, Polak Stefan, Ulicna Marcela, Bohmer Daniel, Vojtassak Jan
Institute of Medical Biology and Genetics, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, Bratislava, Slovak Republic.
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2008 Dec;152(2):235-8. doi: 10.5507/bp.2008.036.
Skeletal muscle contains populations of multipotent adult stem cells also referred to as muscle-derived stem cells.
The main goal of this study was to isolate and culture human adult stem cells from skeletal muscle and characterize them.
Muscle-derived stem cells were isolated from biopsy specimens of femoral muscle. The cells were cultured in Dulbecco's modified Eagle's minimal essential medium supplemented with 10% fetal calf serum and gentamycin. When they reached confluence, they were sub-passaged up to the third passage. Cells from the last passage were prepared for TEM analysis. Production of alpha-actin and desmin was confirmed by histochemistry. Moreover, the phenotypic characterization was performed.
Primary isolated muscle-derived stem cells had a fibroblast-like shape. During subsequent passages they maintained 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. Dilated cisterns of rough endoplasmic reticulum were present in cytoplasm. In certain parts of the cytoplasm there were aggregates of granules of glycogen. The products of cells were actively secreted into the extracellular matrix. They expressed alpha-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 fibroblast surface protein.
Muscle-derived stem cells exhibited typical characteristics typical for mesenchymal stem cells. After analysis of their differentiation potential they could be used in tissue engineering and regenerative medicine.
骨骼肌中含有多能成体干细胞群，也被称为肌肉衍生干细胞。
本研究的主要目标是从骨骼肌中分离并培养人类成体干细胞，并对其进行特性鉴定。
从股肌活检标本中分离肌肉衍生干细胞。细胞在补充有10%胎牛血清和庆大霉素的杜尔贝科改良伊格尔最低限度基本培养基中培养。当细胞达到汇合状态时，进行传代培养，直至第三代。对最后一代细胞进行透射电子显微镜（TEM）分析。通过组织化学确认α-肌动蛋白和结蛋白的产生。此外，进行了表型鉴定。
原代分离的肌肉衍生干细胞呈成纤维细胞样形态。在随后的传代过程中，它们保持了这种形态。TEM分析显示间充质干细胞具有典型的超微结构形态。它们有大的淡染细胞核，含有大量常染色质。细胞核不规则，有明显的核仁。细胞质中存在扩张的粗面内质网池。在细胞质的某些部分有糖原颗粒聚集。细胞产物被积极分泌到细胞外基质中。它们表达α-肌动蛋白和结蛋白。表型鉴定结果表明，几乎所有分析的细胞CD13、CD34、CD56呈阳性，CD45呈阴性。此外，它们不表达抗人成纤维细胞表面蛋白。
肌肉衍生干细胞表现出间充质干细胞的典型特征。在分析其分化潜能后，它们可用于组织工程和再生医学。
