Three-dimensional analysis of the substrate-dependent invasive behavior of a human lung tumor cell line with a confocal laser scanning microscope

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Abstract
Matrigel and collagen G gels were used as models for basement membrane and interstitial space-collagen, respectively, to study the invasive behavior of cells of the human lung tumor cell line EPLC 32M1, which was derived from a squamous cell carcinoma. For three dimensional analysis of the invasive process, cells were seeded onto the gels in a slide chamber and observed with a confocal laser scanning microscope. Optical sectioning in thexy andxz directions and image reconstruction with computer programs allowed us readily to obtain a three-dimensional overview of the invasive process in situ. Both types of gel showed a smooth surface. Matrigel had a granular structure whereas collagen G revealed a fiber-like morphology. The tumor cells showed a matrix-dependent behavior. On Matrigel, within 24 h of incubation, a network of cells appeared on the surface, which developed further within 72 h to interconnected multicellular cords also invading the gel. Tumor cells seeded on collagen G remained individual. They formed pseudopodia and achieved tight contact with the matrix, eventually also invading the gels in a time-dependent manner. Therefore, the composition of the substrate crucially influences the invasion path.