Assessment of the dielectrophoresis (DEP) cross-over frequency (fxo), cell diameter, and derivative membrane capacitance (Cm) 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 fxo and Cm values, the latter of which ranged from 14 to 20 mF/m2. Differentiation of H1 and H9 to a mesenchymal stem cell-like phenotype resulted in similar significant increases in mean Cm values to 41–49 mF/m2 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 Cm value to 28 mF/m2 (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.
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December 2012
Research Article|
December 12 2012
Dielectrophoresis based discrimination of human embryonic stem cells from differentiating derivatives
Srinivas Velugotla;
Srinivas Velugotla
1
Institute for Integrated Micro and Nano Systems, School of Engineering, The University of Edinburgh
, Edinburgh EH9 3JF, United Kingdom
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Steve Pells;
Steve Pells
2
Centre for Regenerative Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
, Chancellor's Building, Edinburgh EH16 4SB, United Kingdom
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Heidi K. Mjoseng;
Heidi K. Mjoseng
2
Centre for Regenerative Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
, Chancellor's Building, Edinburgh EH16 4SB, United Kingdom
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Cairnan R. E. Duffy;
Cairnan R. E. Duffy
2
Centre for Regenerative Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
, Chancellor's Building, Edinburgh EH16 4SB, United Kingdom
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Stewart Smith;
Stewart Smith
1
Institute for Integrated Micro and Nano Systems, School of Engineering, The University of Edinburgh
, Edinburgh EH9 3JF, United Kingdom
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Paul De Sousa;
Paul De Sousa
2
Centre for Regenerative Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
, Chancellor's Building, Edinburgh EH16 4SB, United Kingdom
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Ronald Pethig
Ronald Pethig
1
Institute for Integrated Micro and Nano Systems, School of Engineering, The University of Edinburgh
, Edinburgh EH9 3JF, United Kingdom
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a)
Authors to whom correspondence should be addressed. Electronic addresses: [email protected] and [email protected].
b)
P. De Sousa and R. Pethig contributed equally to this work.
Biomicrofluidics 6, 044113 (2012)
Article history
Received:
August 24 2012
Accepted:
November 27 2012
Citation
Srinivas Velugotla, Steve Pells, Heidi K. Mjoseng, Cairnan R. E. Duffy, Stewart Smith, Paul De Sousa, Ronald Pethig; Dielectrophoresis based discrimination of human embryonic stem cells from differentiating derivatives. Biomicrofluidics 1 December 2012; 6 (4): 044113. https://doi.org/10.1063/1.4771316
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