Cell Therapy and Tissue Engineering Center, Wonju College of Medicine, Yonsei Univ., Wonju, South Korea.
Biochem Biophys Res Commun. 2011 Apr 29;408(1):167-73. doi: 10.1016/j.bbrc.2011.04.002. Epub 2011 Apr 5.
Stem cell therapy for muscular dystrophies requires stem cells that are able to participate in the formation of new muscle fibers. However, the differentiation steps that are the most critical for this process are not clear. We investigated the myogenic phases of human adipose tissue-derived stem cells (hASCs) step by step and the capability of myotube formation according to the differentiation phase by cellular fusion with mouse myoblast C2C12 cells. In hASCs treated with 5-azacytidine and fibroblast growth factor-2 (FGF-2) for 1 day, the early differentiation step to express MyoD and myogenin was induced by FGF-2 treatment for 6 days. Dystrophin and myosin heavy chain (MyHC) expression was induced by hASC conditioned medium in the late differentiation step. Myotubes were observed only in hASCs undergoing the late differentiation step by cellular fusion with C2C12 cells. In contrast, hASCs that were normal or in the early stage were not involved in myotube formation. Our results indicate that stem cells expressing dystrophin and MyHC are more suitable for myotube formation by co-culture with myoblasts than normal or early differentiated stem cells expressing MyoD and myogenin.
用于肌营养不良的干细胞治疗需要能够参与新肌纤维形成的干细胞。然而，对于这个过程最关键的分化步骤尚不清楚。我们逐步研究了人脂肪组织来源的干细胞（hASC）的成肌阶段，并根据与小鼠成肌细胞 C2C12 细胞融合的分化阶段，研究了肌管形成的能力。在经 5-氮杂胞苷和成纤维细胞生长因子-2（FGF-2）处理 1 天后的 hASC 中，通过 FGF-2 处理 6 天诱导早期分化步骤以表达 MyoD 和 myogenin。在晚期分化步骤中，hASC 条件培养基诱导肌球蛋白重链（MyHC）的表达。只有在晚期分化步骤中通过与 C2C12 细胞融合的 hASC 才能观察到肌管。相比之下，正常或早期分化的 hASC 不参与肌管形成。我们的结果表明，表达肌营养不良蛋白和 MyHC 的干细胞比表达 MyoD 和 myogenin 的正常或早期分化的干细胞更适合与成肌细胞共培养形成肌管。
