Department of Chemistry, The University of Western Ontario, London, Ontario, Canada N6A 5B7.
Arch Biochem Biophys. 2012 Dec 1;528(1):21-31. doi: 10.1016/j.abb.2012.08.006. Epub 2012 Aug 25.
Differentiation of pluripotent and lineage restricted stem cells such as neural stem cells (NSCs) was studied on conducting substrates of various nature without perturbation of the genome with exogenous genetic material or chemical stimuli. Primary mouse adult neural stem cells (NSCs) and P19 pluripotent embryonal (P19 EC) carcinoma cells were used. Expression levels of neuronal markers β-III-tubulin and neurofilament were evaluated by immunochemistry and flow cytometry. It was shown that the ability of the substrate to induce differentiation directly correlated with its conductivity. Conducting substrates (conducting oxides or doped π-conjugated organic polymers) with different morphology, structure, and conductivity mechanisms all promoted differentiation of NSC and P19 cells into neuronal lineage to a similar degree without use of additional factors such as poly-L-ornithine coating or retinoic acid, as verified by their morphology and upregulation of the neuronal markers but not astrocyte marker GFAP. However, substrates with low conductance below ca. 10(-4) S cm(-2) did not show this ability. Morphology of differentiating cells was visualized by atomic force microscopy. NSCs cells increased β-III-tubulin expression by 95% and P19 cells by over 30%. Our results suggest that the substrate conductivity is a key factor governing the cell fate. Differentiation of P19 cells into neuronal lineage on conducting substrates was attributed to downregualtion of Akt signaling pathway and increase in expression of dual oxidase 1 (DUOX 1).
在不通过外源遗传物质或化学刺激来干扰基因组的情况下，研究了多能性和谱系受限干细胞（如神经干细胞（NSC））在各种性质的导电基底上的分化。使用了原代成年小鼠神经干细胞（NSC）和 P19 多能胚胎（P19 EC）癌细胞。通过免疫化学和流式细胞术评估神经元标志物 β-III-微管蛋白和神经丝的表达水平。结果表明，基底诱导分化的能力与其导电性直接相关。具有不同形态、结构和导电机理的导电基底（导电氧化物或掺杂的π共轭有机聚合物）都能促进 NSC 和 P19 细胞向神经元谱系分化，程度相似，无需使用额外的因素，如多聚-L-鸟氨酸涂层或维甲酸，这通过其形态和神经元标志物的上调得到验证，但不包括星形胶质细胞标志物 GFAP。然而，导电性低于约 10(-4) S cm(-2)的基底则没有这种能力。通过原子力显微镜观察分化细胞的形态。NSC 细胞的 β-III-微管蛋白表达增加了 95%，P19 细胞增加了 30%以上。我们的结果表明，基底的电导率是决定细胞命运的关键因素。在导电基底上 P19 细胞向神经元谱系的分化归因于 Akt 信号通路的下调和双氧化酶 1（DUOX 1）表达的增加。
