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Biomicrofluidics / Volume 6 / Issue 1 / REGULAR ARTICLES
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Biomicrofluidics 6, 014102 (2012); http://dx.doi.org/10.1063/1.3671399 (14 pages)

Electrotaxis of lung cancer cells in ordered three-dimensional scaffolds

Yung-Shin Sun1, Shih-Wei Peng1,2, Keng-Hui Lin1,3, and Ji-Yen Cheng1,2,4

1Research Center for Applied Sciences, Academia Sinica, Taipei City 11529, Taiwan
2Institute of Biophotonics, National Yang-Ming University, Taipei City 11221, Taiwan
3Institute of Physics, Academia Sinica, Taipei City 11529, Taiwan
4Department of Mechanical and Mechantronic Engineering, National Taiwan Ocean University, Keelung 20224, Taiwan

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(Received 23 September 2011; accepted 30 November 2011; published online 4 January 2012)

In this paper, we report a new method to incorporate 3D scaffold with electrotaxis measurement in the microfluidic device. The electrotactic response of lung cancer cells in the 3D foam scaffolds which resemble the in vivo pulmonary alveoli may give more insight on cellular behaviors in vivo. The 3D scaffold consists of ordered arrays of uniform spherical pores in gelatin. We found that cell morphology in the 3D scaffold was different from that in 2D substrate. Next, we applied a direct current electric field (EF) of 338 mV/mm through the scaffold for the study of cells’ migration within. We measured the migration directedness and speed of different lung cancer cell lines, CL1-0, CL1-5, and A549, and compared with those examined in 2D gelatin-coated and bare substrates. The migration direction is the same for all conditions but there are clear differences in cell morphology, directedness, and migration speed under EF. Our results demonstrate cell migration under EF is different in 2D and 3D environments and possibly due to different cell morphology and/or substrate stiffness.

© 2012 American Institute of Physics

Article Outline

  1. INTRODUCTION
  2. MATERIALS AND METHODS
    1. 3D scaffold fabrication
    2. Fluidic chamber fabrication
    3. EF calculation and measurement
    4. Cell preparation
    5. Electrotaxis experiment
    6. Cell migration measurement
    7. Electrotaxis experiment of lung cancer cells on 2D gelatin substrates
  3. RESULTS
    1. A. 3D scaffolds with a uniform pore size
    2. Comparison of cell morphology in 2D and 3D environments
    3. EF calculation and measurement
    4. Cell migration under dcEF inside 3D scaffolds
    5. Comparison of cell migration under dcEF in 2D and 3D environments
  4. CONCLUSION

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KEYWORDS, PACS, and IPC

Keywords

bioelectric phenomena, biological effects of fields, biological fluid dynamics, biomedical measurement, bioMEMS, cancer, cellular transport, gelatin, lung, microfluidics, substrates

PACS

  • 87.17.Jj

    Cell locomotion, chemotaxis

  • 87.50.cf

    Biophysical mechanisms of interaction

  • 47.61.-k

    Micro- and nano- scale flow phenomena

  • 85.85.+j

    Micro- and nano-electromechanical systems (MEMS/NEMS) and devices

  • 87.80.Ek

    Mechanical and micromechanical techniques

International Patent Classification (IPC)

  • A61B5/00

    Measuring for diagnostic purposes; Identification of persons

  • B81B

    Micro-structural devices or systems, e.g. micro-mechanical devices

  • F15D

    Fluid dynamics, i.e. methods or means for influencing the flow of gases or liquids

ARTICLE DATA

Digital Object Identifier
http://dx.doi.org/10.1063/1.3671399

PUBLICATION DATA

ISSN

1932-1058 (online)

Publisher

$abstract.society.value is a Member of CrossRef American Institute of Physics

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Figures (8) Multimedia (6) Tables (3)

Figures (click on thumbnails to view enlargements)

FIG.1
(a) PDMS-based micro-channel device with solution and gas has been pumped through the inlets and bubbles have been collected from the outlet. (b) Monodisperse bubbles with a uniform size were formed and constantly flowed out.

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FIG.2
Design of the fluidic chamber assembly.

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FIG.3
Side-view of the electrotaxis system.

FIG.3 Download High Resolution Image (.zip file) | Export Figure to PowerPoint

FIG.4
(a) Picture of a 3D scaffold. (b) Bright-field confocal image of a 3D scaffold. Only one fixed layer is shown. Scale bar = 50 μm. (c) Fluorescent confocal image of a 3D scaffold. Only one fixed layer is shown. Scale bar = 50 μm. (d) Re-constructed 3D top-view image of a 3D scaffold. (e) Re-constructed 3D side-view image of a 3D scaffold. (f) Re-constructed 3D image of CL1-0 cells (stained with CellVue®) inside a 3D scaffold (enhanced online) [URL: http://dx.doi.org/10.1063/1.3671399.1 ] .

FIG.4 Download High Resolution Image (.zip file) | Export Figure to PowerPoint

FIG.5
Pulmonary alveoli. An alveolus has a form of a hollow cavity, and in some alveolar walls, there are pores between alveoli called Pores of Kohn (reprinted from The President’s Council on Bioethics, Washington, D.C., January 2009, http://bioethics.georgetown.edu/pcbe/).

FIG.5 Download High Resolution Image (.zip file) | Export Figure to PowerPoint

FIG.6
Ellipticity of (a) CL1-0, (b) CL1-5, and (c) A549 cells after 2 h and 2 days cultured in 2D and 3D environments. (2D: 2D bare substrate, 2DG: 2D-gelatin coated substrate, 3DG: 3D-gelatin made scaffold.) For each cell line, the total number of cells selected for analysis is 30.

FIG.6 Download High Resolution Image (.zip file) | Export Figure to PowerPoint

FIG.7
A549 cells inside a 3D scaffold under an applied EF of 338 mV/mm at (a) t = 0 min and (b) t = 60 min. Clearly some cells (Nos. 1, 2, and 3 in blue circles) migrated through interconnected pores (enhanced online) [URL: http://dx.doi.org/10.1063/1.3671399.2 ] .

FIG.7 Download High Resolution Image (.zip file) | Export Figure to PowerPoint

FIG.8
Polar plots of cell migration after 2 h with and without the applied EF. Left column: CL1-0, CL1-5, and A549 without the applied EF. Right column: CL1-0, CL1-5, and A549 with the applied EF of 338 mV/mm. In the beginning, cells are set at the origin. All plots have the same scale and EF direction (from the right to the left).

FIG.8 Download High Resolution Image (.zip file) | Export Figure to PowerPoint

Multimedia

Tables

Table I. Similarities between 3D porous scaffolds and in vivo pulmonary alveoli.

View Table
Table II. Migration directedness and speed of 3 different cells with and without the applied EF in 3D scaffolds. n is the total number of cells selected for analysis.

View Table
Table III. Electrotaxis of lung cancer cell lines CL1-0, CL1-5, and A549 in 2D and 3D environments. Stimulation time = 2 h (2D: 2D bare substrate, 2DG: 2D-gelatin coated substrate, 3D: 3D-gelatin made scaffold).

View Table


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