Characterization of three-dimensional cancer cell migration in mixed collagen-Matrigel scaffolds using microfluidics and image analysis.
Microfluidic devices are becoming mainstream tools to recapitulate in vitro the behavior of cells and tissues. In this study, we use microfluidic devices filled with hydrogels of mixed collagen-Matrigel composition to study the migration of lung cancer cells under different cancer invasion microenvironments. We present the design of the microfluidic device, characterize the hydrogels morphologically and mechanically and use quantitative image analysis to measure the migration of H1299 lung adenocarcinoma cancer cells in different experimental conditions. Our results show the plasticity of lung cancer cell migration, which turns from mesenchymal in collagen only matrices, to lobopodial in collagen-Matrigel matrices that approximate the interface between a disrupted basement membrane and the underlying connective tissue. Our quantification of migration speed confirms a biphasic role of Matrigel. At low concentration, Matrigel facilitates migration, most probably by providing a supportive and growth factor retaining environment. At high concentration, Matrigel slows down migration, possibly due excessive attachment. Finally, we show that antibody-based integrin blockade promotes a change in migration phenotype from mesenchymal or lobopodial to amoeboid and analyze the effect of this change in migration dynamics, in regards to the structure of the matrix. In summary, we describe and characterize a robust microfluidic platform and a set of software tools that can be used to study lung cancer cell migration under different microenvironments and experimental conditions. This platform could be used in future studies, thus benefitting from the advantages introduced by microfluidic devices: precise control of the environment, excellent optical properties, parallelization for high throughput studies and efficient use of therapeutic drugs.
Laboratory of Preclinical Models and Analytical Tools, Division of Solid Tumors and Biomarkers, Center for Applied Medical Research and CIBERONC, Pamplona, Navarra, Spain.
María
M
Anguiano
Laboratory of Preclinical Models and Analytical Tools, Division of Solid Tumors and Biomarkers, Center for Applied Medical Research and CIBERONC, Pamplona, Navarra, Spain.
Carlos
C
Castilla
Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Brno, Czech Republic.
Martin
M
Maška
Laboratory of Preclinical Models and Analytical Tools, Division of Solid Tumors and Biomarkers, Center for Applied Medical Research and CIBERONC, Pamplona, Navarra, Spain.
Cristina
C
Ederra
Laboratory of Preclinical Models and Analytical Tools, Division of Solid Tumors and Biomarkers, Center for Applied Medical Research and CIBERONC, Pamplona, Navarra, Spain.
Rafael
R
Peláez
Laboratory of Preclinical Models and Analytical Tools, Division of Solid Tumors and Biomarkers, Center for Applied Medical Research and CIBERONC, Pamplona, Navarra, Spain.
Xabier
X
Morales
Biodevices and MEMS group, Water and Health Division, CEIT and TECNUN University of Navarra, Donostia - San Sebastián, Gipuzkoa, SPAIN.
Gorka
G
Muñoz-Arrieta
Biodevices and MEMS group, Water and Health Division, CEIT and TECNUN University of Navarra, Donostia - San Sebastián, Gipuzkoa, SPAIN.
Maite
M
Mujika
Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Brno, Czech Republic.
Michal
M
Kozubek
Bioengineering and Aerospace Engineering Department, Universidad Carlos III de Madrid, Leganes, Madrid.|Biomedical Engineering Division, Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain.
Arrate
A
Muñoz-Barrutia
Department of Biochemistry and Genetics, Faculty of Sciences, University of Navarra, Pamplona, Navarra, Spain.|Department of Immunology and Inmunotherapy, CIMA, Pamplona, Navarra, Spain.
Ana
A
Rouzaut
Biodevices and MEMS group, Water and Health Division, CEIT and TECNUN University of Navarra, Donostia - San Sebastián, Gipuzkoa, SPAIN.
Sergio
S
Arana
Department of Mechanical Engineering, Multiscale in Mechanical and Biological Engineering (M2BE), Aragon Institute of Engineering Research (I3A), University of Zaragoza, Zaragoza, Spain.
José Manuel
JM
Garcia-Aznar
Laboratory of Preclinical Models and Analytical Tools, Division of Solid Tumors and Biomarkers, Center for Applied Medical Research and CIBERONC, Pamplona, Navarra, Spain.
Carlos
C
Ortiz-de-Solorzano
D045744:Cell Line, Tumor; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry / Q000648:ultrastructure; D004058:Diffusion; D004338:Drug Combinations; D005109:Extracellular Matrix; D006801:Humans; D020100:Hydrogels; D007797:Laminin / Q000737:chemistry / Q000648:ultrastructure; D055595:Mechanical Phenomena; D044085:Microfluidics / Q000379:methods; D018613:Microscopy, Confocal; D009362:Neoplasm Metastasis; D010641:Phenotype; D011509:Proteoglycans / Q000737:chemistry / Q000648:ultrastructure; D018874:Spheroids, Cellular; D054457:Tissue Scaffolds / Q000737:chemistry; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment
D004338:Drug Combinations; D020100:Hydrogels; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen