An Automated High-Content Assay for Tumor Cell Migration through 3-Dimensional Matrices

High-content tumor cell migration assays in 3-dimensional (3D) extracellular matrix are a powerful tool for modeling and understanding the biology of this critical step in the process of metastasis. Currently available methods offer very limited throughput and are not amenable to studies of comparative pharmacology or small-scale screening. The authors present an automated approach to high-content tumor cell migration assays. A standard screening-sized plate with an array of microchannels was designed and constructed from common thermoplastics. After filling the channels with 3D matrix, cells were placed at one end of the channel, and migration into the channel was monitored via an imaging system. All liquid-handling steps were performed by standard liquid-handling robotics. Tumor cell migration in the channel was truly 3D and correlated with metastatic potential. The information-rich data from these assays were used to rank the potency of compounds inhibiting migration through 3D collagen as well as to gain additional insights into the compounds’ activities related to cell health. This approach is compatible with a variety of multiparametric, morphological, and/or kinetic readouts.

[1]  Timothy J Mitchison,et al.  Dissecting Temporal and Spatial Control of Cytokinesis with a Myosin II Inhibitor , 2003, Science.

[2]  Melinda Larsen,et al.  Extracellular matrix dynamics in development and regenerative medicine , 2008, Journal of Cell Science.

[3]  M J Bissell,et al.  Tissue architecture and breast cancer: the role of extracellular matrix and steroid hormones. , 2000, Endocrine-related cancer.

[4]  David J Beebe,et al.  A passive pumping method for microfluidic devices. , 2002, Lab on a chip.

[5]  J. Cooper,et al.  Effects of cytochalasin and phalloidin on actin , 1987, The Journal of cell biology.

[6]  I. Fidler,et al.  Metastatic behavior of human tumor cell lines grown in the nude mouse. , 1984, Cancer research.

[7]  R. Bergan,et al.  Genistein Inhibits Matrix Metalloproteinase Type 2 Activation and Prostate Cancer Cell Invasion by Blocking the Transforming Growth Factor β-Mediated Activation of Mitogen-Activated Protein Kinase-Activated Protein Kinase 2-27-kDa Heat Shock Protein Pathway , 2006, Molecular Pharmacology.

[8]  N. Carragher.  Profiling distinct mechanisms of tumour invasion for drug discovery: imaging adhesion, signalling and matrix turnover , 2009, Clinical & Experimental Metastasis.

[9]  T. L. Drell,et al.  Tumor cell locomotion: differential dynamics of spontaneous and induced migration in a 3D collagen matrix. , 2004, Experimental cell research.

[10]  Peter Friedl,et al.  Compensation mechanism in tumor cell migration , 2003, The Journal of cell biology.

[11]  P. Friedl,et al.  Tumour-cell invasion and migration: diversity and escape mechanisms , 2003, Nature Reviews Cancer.

[12]  S. Brenner,et al.  Inhibition of actin polymerization by latrunculin A , 1987, FEBS letters.

[13]  David J Beebe,et al.  Automated cell culture in high density tubeless microfluidic device arrays. , 2008, Lab on a chip.

[14]  Erik Sahai,et al.  Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis , 2003, Nature Cell Biology.

[15]  M J Bissell,et al.  Dynamic reciprocity revisited: a continuous, bidirectional flow of information between cells and the extracellular matrix regulates mammary epithelial cell function. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.