Dielectrophoresis based discrimination of human embryonic stem cells from differentiating derivatives.
Dielectrophoresis based discrimination of human embryonic stem cells from differentiating derivatives.
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DOI：
10.1063/1.4771316
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发表时间：
2012-12
期刊：
影响因子：
3.2
通讯作者：
Srinivas Velugotla;S. Pells;Heidi K. Mjoseng;C. Duffy;Stewart Smith;P. D. De Sousa;R. Pethig
中科院分区：
文献类型：
--
作者：
Srinivas Velugotla;S. Pells;Heidi K. Mjoseng;C. Duffy;Stewart Smith;P. D. De Sousa;R. Pethig
一组未分化的人胚胎干细胞系介电泳（DEP）交叉频率（fxo）、细胞直径和导数膜电容（Cm）值的评估（H1，H9，RCM 1，RH 1），而对于H1的转基因亚克隆（T8），显示hESC系不能根据它们的平均fxo和Cm值来区分，后者的范围为14至20 mF/m（2）。H1和H9分化为间充质干细胞样表型导致两个细胞系的平均Cm值相似地显著增加至41-49 mF/m2（p < 0.0001）。BMP 4诱导的RCM 1向滋养层细胞样表型的分化也导致平均Cm值明显增加至28 mF/m2（p < 0.0001）。在细胞培养物的每次传代后，向更高的膜电容的渐进转变也是明显的，因为H9细胞转变为通过在透明质酸底物上生长诱导的间充质干细胞样状态。这些发现证实了未分化和分化细胞之间存在独特的参数，未来在人胚胎干细胞制造的背景下，介电电泳的应用可以基于这些参数。
Assessment of the dielectrophoresis (DEP) cross-over frequency (f xo), cell diameter, and derivative membrane capacitance (C m) values for a group of undifferentiated human embryonic stem cell (hESC) lines (H1, H9, RCM1, RH1), and for a transgenic subclone of H1 (T8) revealed that hESC lines could not be discriminated on their mean f xo and C m values, the latter of which ranged from 14 to 20 mF/m(2). Differentiation of H1 and H9 to a mesenchymal stem cell-like phenotype resulted in similar significant increases in mean C m values to 41-49 mF/m(2) in both lines (p < 0.0001). BMP4-induced differentiation of RCM1 to a trophoblast cell-like phenotype also resulted in a distinct and significant increase in mean C m value to 28 mF/m(2) (p < 0.0001). The progressive transition to a higher membrane capacitance was also evident after each passage of cell culture as H9 cells transitioned to a mesenchymal stem cell-like state induced by growth on a substrate of hyaluronan. These findings confirm the existence of distinctive parameters between undifferentiated and differentiating cells on which future application of dielectrophoresis in the context of hESC manufacturing can be based.