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Biomedical Microdevices

, Volume 16, Issue 3, pp 465–478 | Cite as

The F-actin and adherence-dependent mechanical differentiation of normal epithelial cells after TGF-β1-induced EMT (tEMT) using a microplate measurement system

  • T. H. Wu
  • Y. W. Chiou
  • W. T. Chiu
  • M. J. Tang
  • C. H. Chen
  • Ming-Long YehEmail author
Article

Abstract

The epithelial to mesenchymal transition (EMT) is known to involve several physiological and pathological phenomena. In this study, we utilized a microplate measurement system (MMS) approach based on the deflection of a flexible micro-cantilever to measure cell stiffness (in Pa) and adhesion force (in nN) of a single cell during EMT with nN resolution. Our results demonstrated that after transforming growth factor-β1 (TGF-β1) induced EMT (tEMT), NMuMG cells became stiffer due to thicker and more abundant F-actin and displayed stronger vinculin accumulation after long-term cell-substrate adhesion. The MMS could distinguish differences in compressive stiffness (219 ± 10 and 287 ± 14 Pa), tensile stiffness (114 ± 14 and 132 ± 12 Pa), and adhesion force (150 ± 42 and 192 ± 31 nN) between cells before and after tEMT. However, without proper development of the F-actin structure and adequate adherent time, the mechanical differences were diminished. After tEMT, the cells with increased stiffness and a cell-substrate adhesion force benefited by migrating more rapidly and had more invasiveness. Thus, this technology has the potential to benefit research focused on cancer diagnosis, drug development, and cell-substrate interactions.

Keywords

Epithelial to mesenchymal transitions (EMT) Normal marine mammary gland cells (NMuMG) Transforming growth factor-β1 (TGF-β1) Atomic force microscopy (AFM) Cell stiffness Adhesion force 

Notes

Acknowledgments

The authors acknowledge financial support from the National Science Council of Taiwan (98-2627-B-006-009-, 99-2627-B-006-009-, 100-2627-B-006-009-).

Supplementary material

10544_2014_9849_MOESM1_ESM.docx (348 kb)
Figure S1 (DOCX 347 kb)
10544_2014_9849_MOESM2_ESM.docx (736 kb)
Figure S2 (DOCX 736 kb)
Supplementary Video 1

(AVI 24023 kb)

Supplementary Video 2

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10544_2014_9849_MOESM5_ESM.docx (22 kb)
Supplementary Table 1 (DOCX 22 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • T. H. Wu
    • 1
  • Y. W. Chiou
    • 1
  • W. T. Chiu
    • 1
  • M. J. Tang
    • 2
  • C. H. Chen
    • 3
  • Ming-Long Yeh
    • 1
    Email author
  1. 1.Department of Biomedical EngineeringNational Cheng Kung UniversityTainan CityTaiwan
  2. 2.Institute of PhysiologyNational Cheng Kung UniversityTainanTaiwan
  3. 3.Department of RehabilitationKaohsiung Municipal Ta-Tung HospitalKaohsiungTaiwan

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