Modulating the Cellular Mechanics by Regulating the Cytoskeletal Proteins

The goal of this project was to study the effect of low vimentin levels on the structure, mechanics, and migration/invasiveness of cells. To do this, we performed immunocytochemistry, confocal imaging, atomic force microscopy, and wound healing assays on NIH 3T3 cells that had vimentin knocked down by shRNA to characterize vimentin’s effects. We also worked to develop an invasion assay that better recreates in vivo conditions and could be better observed for any invasive cell types. An assay was developed, which utilized a microfluidic device that utilizes a Matrigel barrier flanked by a cell culture chamber and a chamber containing a chemoattractant of choice. Our system, if refined, would allow direct observation and quantification of cell invasiveness both in the 2D and 3D mode