Institute for Integrated Micro and Nano Systems, School of Engineering, The University of Edinburgh, Edinburgh EH9 3JF, United Kingdom.
Biomicrofluidics. 2012 Aug 13;6(3):34113. doi: 10.1063/1.4746252. Print 2012 Sep.
Myoblasts are muscle derived mesenchymal stem cell progenitors that have great potential for use in regenerative medicine, especially for cardiomyogenesis grafts and intracardiac cell transplantation. To utilise such cells for pre-clinical and clinical applications, and especially for personalized medicine, it is essential to generate a synchronised, homogenous, population of cells that display phenotypic and genotypic homogeneity within a population of cells. We demonstrate that the biomarker-free technique of dielectrophoresis (DEP) can be used to discriminate cells between stages of differentiation in the C2C12 myoblast multipotent mouse model. Terminally differentiated myotubes were separated from C2C12 myoblasts to better than 96% purity, a result validated by flow cytometry and Western blotting. To determine the extent to which cell membrane capacitance, rather than cell size, determined the DEP response of a cell, C2C12 myoblasts were co-cultured with GFP-expressing MRC-5 fibroblasts of comparable size distributions (mean diameter ∼10 μm). A DEP sorting efficiency greater than 98% was achieved for these two cell types, a result concluded to arise from the fibroblasts possessing a larger membrane capacitance than the myoblasts. It is currently assumed that differences in membrane capacitance primarily reflect differences in the extent of folding or surface features of the membrane. However, our finding by Raman spectroscopy that the fibroblast membranes contained a smaller proportion of saturated lipids than those of the myoblasts suggests that the membrane chemistry should also be taken into account.
成肌细胞是一种具有巨大潜力的肌肉来源间充质干细胞祖细胞，可用于再生医学，特别是用于心肌发生移植物和心脏内细胞移植。为了将这些细胞用于临床前和临床应用，特别是用于个性化医学，必须生成同步、同质的细胞群体，该群体在细胞群体中显示出表型和基因型的均一性。我们证明，无生物标志物的电介质电泳 (DEP) 技术可用于区分 C2C12 成肌细胞多能鼠模型中分化阶段的细胞。终末分化的肌管与 C2C12 成肌细胞分离，纯度优于 96%，这一结果通过流式细胞术和 Western blot 得到验证。为了确定细胞膜电容而不是细胞大小决定细胞 DEP 反应的程度，将 C2C12 成肌细胞与 GFP 表达的大小分布相似的 MRC-5 成纤维细胞共培养（平均直径约为 10 μm）。这两种细胞类型的 DEP 分选效率均大于 98%，这一结果可以归因于成纤维细胞具有比成肌细胞更大的细胞膜电容。目前假设，细胞膜电容的差异主要反映了细胞膜折叠程度或表面特征的差异。然而，我们通过拉曼光谱发现，成纤维细胞的膜中含有较少比例的饱和脂质，而肌细胞中含有较多的饱和脂质，这表明膜化学也应该被考虑在内。
