Label‐free profiling of cell dynamics: A sequence of impedance‐based assays to estimate tumor cell invasiveness in vitro

&NA; Dynamic properties of cancer cells, most notably their ability to migrate, have been correlated successfully with their invasive nature in vivo. To establish a stronger experimental basis for such a correlation we subjected five different cancer cell lines of well‐defined metastatic potential to a sequence of three independent assays reporting on three different aspects of cell dynamics, namely (1) the kinetics of cell spreading, (2) cell shape fluctuations, and (3) cell migration. The sequentially applied assays correspond to different measuring modes of the well‐established ECIS technique that is based on non‐invasive and label‐free impedance readings of planar gold‐film electrodes that serve as the growth substrate for the cells under study. Every individual assay returned a characteristic parameter describing the behavior of the cell lines in that particular assay quantitatively. The parameters of all three assays were ranked to establish individual profiles of cell dynamics for every cell line that correlate favorably with the cells' invasive properties. The sequence of impedance‐based assays described here requires only small cell populations (< 10.000 cells), it is highly automated and easily adapted to 96‐well formats. It provides an in‐depth dynamic profile of adherent cells that might be useful in other areas besides cancer research as well. Graphical abstract Figure. No caption available. HighlightsThree impedance‐based assays were applied sequentially to one given cell population.Assays report on (a) cell adhesion, (b) cell motility and (c) collective cell migration.Results provide a profile of cell dynamics.Five cancer cell lines with well‐known metastatic potential were studied.In vitro profile of cell dynamics correlates with metastatic potential in vivo.

[1]  M. Herlyn.  Human melanoma: Development and progression , 1990, Cancer and Metastasis Reviews.

[2]  Adam J Engler,et al.  Metastatic State of Cancer Cells May Be Indicated by Adhesion Strength. , 2017, Biophysical journal.

[3]  K. Syrigos,et al.  Cell Adhesion Molecules: Role and Clinical Significance in Cancer , 2009, Cancer investigation.

[4]  T. Mitchison,et al.  A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods , 2004, BMC biotechnology.

[5]  J. Jankowski,et al.  Alterations in cadherin and catenin expression during the biological progression of melanocytic tumours. , 1999, Molecular pathology : MP.

[6]  Judith P. Johnson.  Cell Adhesion Molecules in the Development and Progression of Malignant Melanoma , 2004, Cancer and Metastasis Reviews.

[7]  J. Wegener,et al.  Cell Growth and Cell Death Studied by Electric Cell-Substrate Impedance Sensing , 2012 .

[8]  Joachim Wegener,et al.  Electrical wound-healing assay for cells in vitro. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[9]  G. Röderer,et al.  Toxicological investigations on the respirable fraction of silicon carbide grain products by the in vitro vector model , 2014, Inhalation toxicology.

[10]  Gabriel Fenteany,et al.  Signaling pathways and cell mechanics involved in wound closure by epithelial cell sheets , 2000, Current Biology.

[11]  J. Wegener,et al.  Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. , 2000, Experimental cell research.

[12]  L. Puricelli,et al.  Modulation of fibronectin expression and proteolytic activity associated with the invasive and metastatic phenotype in two new murine mammary tumor cell lines. , 1997, International journal of oncology.

[13]  I. Giaever,et al.  Micromotion of mammalian cells measured electrically. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Andreas Janshoff,et al.  Cell motility probed by noise analysis of thickness shear mode resonators. , 2006, Analytical chemistry.

[15]  C. Lo,et al.  pH changes in pulsed CO2 incubators cause periodic changes in cell morphology. , 1994, Experimental cell research.

[16]  C. Lo,et al.  Distinguishing cancerous from noncancerous cells through analysis of electrical noise. , 2007, Physical review. E, Statistical, nonlinear, and soft matter physics.

[17]  C. Lo,et al.  Correlated Motion and Oscillation of Neighboring Cells in Vitro , 2001, Cell communication & adhesion.

[18]  A. Lityńska,et al.  Expression of beta1-integrins and N-cadherin in bladder cancer and melanoma cell lines. , 2000, Acta biochimica Polonica.

[19]  Joachim Wegener,et al.  Real-time impedance assay to follow the invasive activities of metastatic cells in culture. , 2002, BioTechniques.

[20]  A. Üren,et al.  A real-time electrical impedance based technique to measure invasion of endothelial cell monolayer by cancer cells. , 2011, Journal of visualized experiments : JoVE.

[21]  P. Forscher,et al.  A novel cytoskeletal structure involved in purse string wound closure and cell polarity maintenance , 1993, The Journal of cell biology.

[22]  I. Giaever,et al.  Monitoring fibroblast behavior in tissue culture with an applied electric field. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[23]  S. Galli,et al.  Characterization of a fibroblastoid mammary carcinoma cell line (LM2) originated from a mouse adenocarcinoma. , 2000, International journal of oncology.

[24]  J. Tímár,et al.  Two human melanoma xenografts with different metastatic capacity and glycosaminoglycan pattern , 2004, Journal of Cancer Research and Clinical Oncology.

[25]  J. Stockman,et al.  Distinct Sets of Genetic Alterations in Melanoma , 2007 .

[26]  C. Liang,et al.  In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro , 2007, Nature Protocols.

[27]  Andreas Janshoff,et al.  Dynamics of human cancer cell lines monitored by electrical and acoustic fluctuation analysis. , 2010, Integrative biology : quantitative biosciences from nano to macro.

[28]  C. Lo,et al.  Monitoring motion of confluent cells in tissue culture. , 1993, Experimental cell research.

[29]  J. Tímár,et al.  Selection and characterization of human melanoma lines with different liver‐colonizing capacity , 1990, International journal of cancer.

[30]  S. Mok,et al.  Monitoring of ovarian cancer cell invasion in real time with frequency-dependent impedance measurement. , 2016, American journal of physiology. Cell physiology.

[31]  Chun-Min Lo,et al.  Detecting effects of low levels of cytochalasin B in 3T3 fibroblast cultures by analysis of electrical noise obtained from cellular micromotion. , 2009, Biosensors & bioelectronics.

[32]  O. Thoumine,et al.  Predicting the kinetics of cell spreading. , 2002, Journal of biomechanics.