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Impedance-based real-time monitoring of neural stem cell differentiation

F. J. Shah

F. J. Shah

  • Department of Micro- and Nanotechnology, Technical University of DenmarkKongens Lyngby, Denmark
  • Particle Analytical ApSørsholm, Denmark
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Shah, F. J.
, 
C. Caviglia

C. Caviglia

  • Department of Micro- and Nanotechnology, Technical University of DenmarkKongens Lyngby, Denmark
  • Radiometer Medical ApSønshøj, Denmark
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Caviglia, C.
, 
K. Zór

K. Zór

  • Department of Micro- and Nanotechnology, Technical University of DenmarkKongens Lyngby, Denmark
  • Center for Intelligent Drug Delivery and Sensing Using Microcontainers and Nanomechanics, Department of Health Technology, Technical University of DenmarkKongens Lyngby, Denmark
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Zór, K.
, 
M. Carminati

M. Carminati

Dipartimento di Elettronica, Informazione e Bioingegneria - DEIB, Politecnico di MilanoMilano, Italy
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Carminati, M.
, 
G. Ferrari

G. Ferrari

Dipartimento di Elettronica, Informazione e Bioingegneria - DEIB, Politecnico di MilanoMilano, Italy
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Ferrari, G.
, 
M. Sampietro

M. Sampietro

Dipartimento di Elettronica, Informazione e Bioingegneria - DEIB, Politecnico di MilanoMilano, Italy
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Sampietro, M.
, 
A. Martínez-Serrano

A. Martínez-Serrano

Department of Molecular Neuropathology, Center of Molecular Biology Severo Ochoa, Universidad Autónoma de MadridMadrid, Spain
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Martínez-Serrano, A.
, 
J. K. Emnéus

J. K. Emnéus

  • Department of Micro- and Nanotechnology, Technical University of DenmarkKongens Lyngby, Denmark
  • Present affiliation: Department of Biotechnology and Biomedicine, Technical University of DenmarkKongens Lyngby, Denmark
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Emnéus, J. K.
 y 
A. R. Heiskanen

A. R. Heiskanen

  • Department of Micro- and Nanotechnology, Technical University of DenmarkKongens Lyngby, Denmark
  • Present affiliation: Department of Biotechnology and Biomedicine, Technical University of DenmarkKongens Lyngby, Denmark
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Heiskanen, A. R.
  
06 oct 2021
Impedance-based real-time monitoring of neural stem cell differentiation's Cover Image
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Publicado en línea: 06 oct 2021
Páginas: 34 - 49
Recibido: 23 dic 2020
DOI: https://doi.org/10.2478/joeb-2021-0006
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© 2021 Shah F. J., Caviglia C., Zór K., Carminati M., Ferrari G., Sampietro M., Martínez-Serrano A., Emnéus J. K., Heiskanen A. R., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1

Impedance measurement setup: A) Microelectrode array chip with 12 IDEs; B) zoom-in view of one IDE (the ca. 500 μm x 500 μm opening in the silicon nitride passivation layer appears as a lighter region in the center) ; C) chip holder (the lower plate accommodates a MEA chip and the upper plate provides the 600 μl cell culture chamber and an array of holes for electrical connections using spring-loaded pins; fluid tight sealing on the MEA chip is achieved by using a laser cut silicon rubber gasket); D) Printed circuit board (PCB) of the custom-made 12-channel bipotentiostat (the PCB has an opening to the cell culture vial of the chip holder to allow liquid handling and microscopic visualization; E) user interface of the data acquisition software showing recorded impedance magnitude vs. log frequency.
Impedance measurement setup: A) Microelectrode array chip with 12 IDEs; B) zoom-in view of one IDE (the ca. 500 μm x 500 μm opening in the silicon nitride passivation layer appears as a lighter region in the center) ; C) chip holder (the lower plate accommodates a MEA chip and the upper plate provides the 600 μl cell culture chamber and an array of holes for electrical connections using spring-loaded pins; fluid tight sealing on the MEA chip is achieved by using a laser cut silicon rubber gasket); D) Printed circuit board (PCB) of the custom-made 12-channel bipotentiostat (the PCB has an opening to the cell culture vial of the chip holder to allow liquid handling and microscopic visualization; E) user interface of the data acquisition software showing recorded impedance magnitude vs. log frequency.

Figure 2

Cell Index vs. time for LUHMES cells: Initial cell density (cells/cm2) A) 30,000; B) 60,000; C) 120,000. Proliferating (blue) and differentiating (red) cells. Time in days after cell seeding. (Error bars: s.e.m., n = 6).
Cell Index vs. time for LUHMES cells: Initial cell density (cells/cm2) A) 30,000; B) 60,000; C) 120,000. Proliferating (blue) and differentiating (red) cells. Time in days after cell seeding. (Error bars: s.e.m., n = 6).

Figure 3

Fluorescence microscopy images of live stained (Calcein AM) LUHMES cells in growth medium (GM) and differentiation medium (DM). Initial cell density 60,000 cells/cm2. Time in days after cell seeding. (Scale bars: 50 μm).
Fluorescence microscopy images of live stained (Calcein AM) LUHMES cells in growth medium (GM) and differentiation medium (DM). Initial cell density 60,000 cells/cm2. Time in days after cell seeding. (Scale bars: 50 μm).

Figure 4

Cell Index vs. time for hVM1 Bcl-XL cells: Initial cell density 120,000 cells/cm2. Proliferating (blue) and differentiating (red) cells. Time in days after cell seeding. (Error bars: s.e.m., n = 6).
Cell Index vs. time for hVM1 Bcl-XL cells: Initial cell density 120,000 cells/cm2. Proliferating (blue) and differentiating (red) cells. Time in days after cell seeding. (Error bars: s.e.m., n = 6).

Figure 5

Fluorescence microscopy images of live stained (Calcein AM) hVM1 Bcl-XL cells in growth medium (GM) and differentiation medium (DM). Initial cell density 120,000 cells/cm2. Time in days after cell seeding. (Scale bars: 50 μm)
Fluorescence microscopy images of live stained (Calcein AM) hVM1 Bcl-XL cells in growth medium (GM) and differentiation medium (DM). Initial cell density 120,000 cells/cm2. Time in days after cell seeding. (Scale bars: 50 μm)

Figure 6

Equivalent circuit models for analysis of impedance spectra acquired A) in the presence and B) in the absence of cells. (Cell specific parameters: Rextra, Rcell, Ccell). For detailed description of the components, see the text.
Equivalent circuit models for analysis of impedance spectra acquired A) in the presence and B) in the absence of cells. (Cell specific parameters: Rextra, Rcell, Ccell). For detailed description of the components, see the text.

Figure 7

Example of typical Bode plots for an electrode and the same electrode 48 h after seeding of 60,000 LUHMES cells/cm2: A) impedance magnitude and B) phase angle. Solid lines show the nonlinear least squares fit of the experimental data to the equivalent circuit models of Fig. 6.
Example of typical Bode plots for an electrode and the same electrode 48 h after seeding of 60,000 LUHMES cells/cm2: A) impedance magnitude and B) phase angle. Solid lines show the nonlinear least squares fit of the experimental data to the equivalent circuit models of Fig. 6.

Figure 8

Summary of the cell specific equivalent circuit components (Rcell, Rextra, Ccell) for A) proliferating and B) differentiating LUHMES cells (seeding density: 60,000 cells/cm2). Time in days after cell seeding. (Error bars: s.e.m., n = 6).
Summary of the cell specific equivalent circuit components (Rcell, Rextra, Ccell) for A) proliferating and B) differentiating LUHMES cells (seeding density: 60,000 cells/cm2). Time in days after cell seeding. (Error bars: s.e.m., n = 6).

Figure 9

Summary of the cell specific equivalent circuit components (Rcell, Rextra, Ccell) for A) proliferating and B) differentiating hVM1 Bcl-XL cells (seeding density: 120,000 cells/cm2). Time in days after cell seeding. (Error bars: s.e.m., n = 6).
Summary of the cell specific equivalent circuit components (Rcell, Rextra, Ccell) for A) proliferating and B) differentiating hVM1 Bcl-XL cells (seeding density: 120,000 cells/cm2). Time in days after cell seeding. (Error bars: s.e.m., n = 6).

Figure S1

Normalized impedance vs. frequency for A) LUHMES (60,000 cells/cm2) and B) hVM1 Bcl-XL (120,000 cells/cm2). The spectra were acquired 48 h after cell seeding. (Error bars: standard deviation, n = 3)
Normalized impedance vs. frequency for A) LUHMES (60,000 cells/cm2) and B) hVM1 Bcl-XL (120,000 cells/cm2). The spectra were acquired 48 h after cell seeding. (Error bars: standard deviation, n = 3)

Figure S2

Cell Index vs. time for proliferating hVM1 Bcl-XL cells. Initial cell seeding density 60,000 cells/cm2 and 120,000 cells/cm2. (Error bars: standard deviation, n = 3)
Cell Index vs. time for proliferating hVM1 Bcl-XL cells. Initial cell seeding density 60,000 cells/cm2 and 120,000 cells/cm2. (Error bars: standard deviation, n = 3)
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Ingeniería, Bioingeniería e ingeniería biomédica, Electrónica biomédica, Ciencias de la vida, Biofísica, Medicina, Ingeniería biomédica, Física, Espectroscopia y metrología
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