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Impedimetric melanoma invasion assay device using a simple paper membrane and stencil-printed electrode on PMMA substrate

Pupinyo, Naricha ; Heiskanen, Arto LU ; Chailapakul, Orawon ; Gorton, Lo LU ; Emnéus, Jenny LU and Laiwattanapaisal, Wanida (2020) In Sensing and Bio-Sensing Research 29.
Abstract

The transwell assay is currently the most popular approach to studying cellular invasion due to its ease of use and readout, and the possibility for quantitative measurements. However, it only allows end-point measurements without the possibility for real-time tracking of the dynamics of cell movement during an invasion. Moreover, it requires cell labeling, and construction of customized devices is hampered by the commercial standard membrane inserts, only available in certain designs. Recently, paper has been used as a scaffold for three-dimensional (3D) cell cultures. Because of its microfibrous structure and easy handling, it could be a versatile alternative as a membrane insert in customized devices. Here, we develop a low-cost... (More)

The transwell assay is currently the most popular approach to studying cellular invasion due to its ease of use and readout, and the possibility for quantitative measurements. However, it only allows end-point measurements without the possibility for real-time tracking of the dynamics of cell movement during an invasion. Moreover, it requires cell labeling, and construction of customized devices is hampered by the commercial standard membrane inserts, only available in certain designs. Recently, paper has been used as a scaffold for three-dimensional (3D) cell cultures. Because of its microfibrous structure and easy handling, it could be a versatile alternative as a membrane insert in customized devices. Here, we develop a low-cost real-time invasion assay device using paper as an alternative membrane insert. The device was designed for two-electrode impedance measurements and fabricated using CNC micromilling. It also comprised a disposable low-cost stencil-printed working electrode on a poly(methyl methacrylate) substrate below the membrane and glassy carbon counter electrode above the membrane inserted in a specially designed lid. Thus, the impedance measurements during cell invasion addressed the entire membrane. We demonstrated the function of the device by monitoring the invasion of B16 melanoma 4A5 cells from a mouse using insulin growth factor-1 as the chemoattractant. The cell invasion on paper was visualized using scanning electron microscopy and confocal microscopy with Z-stack 3D imaging. Melanoma cell invasion could be observed within 7 h after the chemoattractant treatment, which was faster than the conventional assay and less likely to be influenced by cell proliferation.

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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
3D cell culture, Impedance monitoring, Invasion assay, Melanoma, Paper-based cell culture, Stencil-printed electrode
in
Sensing and Bio-Sensing Research
volume
29
article number
100354
publisher
Elsevier
external identifiers
  • scopus:85085375586
ISSN
2214-1804
DOI
10.1016/j.sbsr.2020.100354
language
English
LU publication?
yes
id
9487150d-86a1-4c20-85d1-f3f6c47256c0
date added to LUP
2020-06-12 12:11:15
date last changed
2020-08-13 16:36:54
@article{9487150d-86a1-4c20-85d1-f3f6c47256c0,
  abstract     = {<p>The transwell assay is currently the most popular approach to studying cellular invasion due to its ease of use and readout, and the possibility for quantitative measurements. However, it only allows end-point measurements without the possibility for real-time tracking of the dynamics of cell movement during an invasion. Moreover, it requires cell labeling, and construction of customized devices is hampered by the commercial standard membrane inserts, only available in certain designs. Recently, paper has been used as a scaffold for three-dimensional (3D) cell cultures. Because of its microfibrous structure and easy handling, it could be a versatile alternative as a membrane insert in customized devices. Here, we develop a low-cost real-time invasion assay device using paper as an alternative membrane insert. The device was designed for two-electrode impedance measurements and fabricated using CNC micromilling. It also comprised a disposable low-cost stencil-printed working electrode on a poly(methyl methacrylate) substrate below the membrane and glassy carbon counter electrode above the membrane inserted in a specially designed lid. Thus, the impedance measurements during cell invasion addressed the entire membrane. We demonstrated the function of the device by monitoring the invasion of B16 melanoma 4A5 cells from a mouse using insulin growth factor-1 as the chemoattractant. The cell invasion on paper was visualized using scanning electron microscopy and confocal microscopy with Z-stack 3D imaging. Melanoma cell invasion could be observed within 7 h after the chemoattractant treatment, which was faster than the conventional assay and less likely to be influenced by cell proliferation.</p>},
  author       = {Pupinyo, Naricha and Heiskanen, Arto and Chailapakul, Orawon and Gorton, Lo and Emnéus, Jenny and Laiwattanapaisal, Wanida},
  issn         = {2214-1804},
  language     = {eng},
  publisher    = {Elsevier},
  series       = {Sensing and Bio-Sensing Research},
  title        = {Impedimetric melanoma invasion assay device using a simple paper membrane and stencil-printed electrode on PMMA substrate},
  url          = {http://dx.doi.org/10.1016/j.sbsr.2020.100354},
  doi          = {10.1016/j.sbsr.2020.100354},
  volume       = {29},
  year         = {2020},
}