Invadopodia formation assay and quantification analysis with confocal system in A549 cells.

<p>The cells in control group (A), EGF group (B), and GM6001/EGF group (C), were cultured on a 3D microfluidic device for 12 h. The cells were stained green represented combined with cortactin, stained red represented combined with F-actin, and arrowheads in merge pictures indicated cells displaying invadopodia. (D) The percent of the cells with invadopodia formation. (E) The average number of invadopodia per cell. Error bars represented the SD of three different determinations. *Statistically significant between control group and EGF group; **statistically significant between EGF group and GM6001/EGF group, p<0.05. Magnification: ×1200.</p

Actual invadopodia formation of A549 cells in control group (A), EGF group (B), and GM6001/EGF group (C) in 3D extracellular matrix in the microfluidic device with confocal system.

<p>Invadopodia could be obviously induced by EGF in (B), <b>while</b> this induction could be inhibited by GM6001 in (C). White arrowheads represented invadopodia. Magnification: ×1200.</p

Fluoresent analysis of apoptotic in A549 cells.

<p>Cells were cultured on the 3D microfluidic device for 96 h and then stained by Hoechst and PI. Live cells were stained blue and dead cells were stained red. Magnification: ×200.</p

Illustration of medium flow direction in the microfluidic device.

<p>The blue, red and green indicators represented control group, EGF group, GM6001/EGF group respectively. These three indicators were perfused into microchannels from inlet A, B, C simultaneously and separately, while these indicators could spread out to cell chambers of both sides via oval microchannels uniformly and in parallel without crossing.</p

A microfluidic chip designed for the study of cancer cells invasion in 3D matrix.

<p>(A) Schematic representation of the microfluidic platform. Layout of the integrated microfluidic device is composed of three units sharing a common outlet, each of which contains an inlet, three parallel main channels, three cell culture chambers and an outlet. (B) A magnified illustration of one cell culture chamber. (C) Photograph of the microfluidic system.</p