Pang R, Zhang Y, Pan X, Gu R, Hou X, Xiang P, Liu Z, Zhu X, Hu J, Zhao J, Zhang C
Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, P.R. China.
Cell Mol Biol (Noisy-le-grand). 2010 Apr 15;56 Suppl:OL1276-85.
Embryonic-like stem cell (ELSC), expressing part of surface markers of human embryonic stem cells, may be a better candidate for cell therapy of degenerative muscular disease than mesenchymal stem cell (MSC). We isolated ELSC and MSC from bone marrow, respectively, and compared their differences in the characteristics and the capacity of myogenic differentiation. Results showed that ELSC could be isolated successfully from 3 adult bone marrow samples by using serum-free medium with 10ng/ml basic fibroblast growth factor (bFGF). At the same cell density, MSC could also be isolated from the same samples by using DMEM/F12 medium containing 10% new cattle serum. However, ELSC appeared as small, morphologically slenderer, upregulated expression of SSEA-4 and ultramicroscopically more immature than MSC derived from the same samples. Immunofluorescent staining and RT-PCR analysis showed ELSC weakly expressed Oct-4, Nanog-3 and Sox-2. Moreover, ELSC and MSC could be induced into long, multinucleated fibers expressing myogenin and myosin heavy chain (MHC) in myogenic differentiation medium, but by day 10, proportion of multinucleated fibers positive for MHC was respectively 25.0%+/-6.9% and 13.8%+/-7.6% in ELSC and MSC culture. These data suggest that bone marrow derived ELSC represent an ideal candidate for cell therapy of degenerative muscular disease.
胚胎样干细胞(ELSC)表达人胚胎干细胞的部分表面标志物,与间充质干细胞(MSC)相比,可能是退行性肌肉疾病细胞治疗的更好候选者。我们分别从骨髓中分离出ELSC和MSC,并比较了它们在特性和肌源性分化能力方面的差异。结果表明,通过使用含有10ng/ml碱性成纤维细胞生长因子(bFGF)的无血清培养基,可成功从3份成人骨髓样本中分离出ELSC。在相同细胞密度下,使用含有10%新生牛血清的DMEM/F12培养基也可从相同样本中分离出MSC。然而,与来自相同样本的MSC相比,ELSC体积较小,形态更细长,SSEA-4表达上调,超微结构上更不成熟。免疫荧光染色和RT-PCR分析显示ELSC弱表达Oct-4、Nanog-3和Sox-2。此外,在肌源性分化培养基中,ELSC和MSC均可被诱导形成表达生肌调节因子和肌球蛋白重链(MHC)的长的多核纤维,但在第10天时,ELSC和MSC培养物中MHC阳性的多核纤维比例分别为25.0%±6.9%和13.8%±7.6%。这些数据表明,骨髓来源的ELSC是退行性肌肉疾病细胞治疗的理想候选者。
Cytotherapy. 2013-1-9
Stem Cells Dev. 2010-10-29