Caddick Jenny, Kingham Paul J, Gardiner Natalie J, Wiberg Mikael, Terenghi Giorgio
Blond McIndoe Research Laboratories, University of Manchester, Manchester, United Kingdom.
Glia. 2006 Dec;54(8):840-9. doi: 10.1002/glia.20421.
We have investigated the phenotypic and bioassay characteristics of bone marrow mesenchymal stromal cells (MSCs) differentiated along a Schwann cell lineage using glial growth factor. Expression of the Schwann cell markers S100, P75, and GFAP was determined by immunocytochemical staining and Western blotting. The levels of the stem cell markers Stro-1 and alkaline phosphatase and the neural progenitor marker nestin were also examined throughout the differentiation process. The phenotypic properties of cells differentiated at different passages were also compared. In addition to a phenotypic characterization, the functional ability of differentiated MSCs has been investigated employing a co-culture bioassay with dissociated primary sensory neurons. Following differentiation, MSCs underwent morphological changes similar to those of cultured Schwann cells and stained positively for all three Schwann cell markers. Quantitative Western blot analysis showed that the levels of S100 and P75 protein were significantly elevated upon differentiation. Differentiated MSCs were also found to enhance neurite outgrowth in co-culture with sensory neurons to a level equivalent or superior to that produced by Schwann cells. These findings support the assertion that MSCs can be differentiated into cells that are Schwann cell-like in terms of both phenotype and function.
我们利用胶质生长因子研究了沿雪旺细胞谱系分化的骨髓间充质基质细胞(MSCs)的表型和生物测定特征。通过免疫细胞化学染色和蛋白质印迹法测定雪旺细胞标志物S100、P75和GFAP的表达。在整个分化过程中还检测了干细胞标志物Stro-1和碱性磷酸酶以及神经祖细胞标志物巢蛋白的水平。还比较了不同传代分化细胞的表型特性。除了表型特征外,还采用与解离的初级感觉神经元共培养生物测定法研究了分化的MSCs的功能能力。分化后,MSCs发生了与培养的雪旺细胞相似的形态变化,并且对所有三种雪旺细胞标志物染色均呈阳性。蛋白质印迹定量分析表明,分化后S100和P75蛋白水平显著升高。还发现分化的MSCs与感觉神经元共培养时可增强神经突生长,达到与雪旺细胞相当或更高的水平。这些发现支持了MSCs可以分化为在表型和功能上都类似雪旺细胞的细胞这一论断。