Cell motility plays an important role in development, wound healing and cancer progression. A fundamental unresolved challenge in the field is to obtain reliable measures of motility metrics from single cells and then derive statistically meaningful data on cell population level motility behavior. Currently available tools are limited, for instance, they track cells as unrelated objects (i.e., do not consider cell division), lack ability for high-throughput dynamic parameter extraction, or employ inaccurate tracking algorithms. To extract dynamic morphology and motility parameters at the single cell level we have developed CellAnimation, an open-source high-throughput microscopy framework written in MATLAB which is currently being used in several labs at Vanderbilt and elsewhere. We have also developed a novel cell tracking algorithm which supports mitotic event detection and ancestry recording and we have shown that it outperforms the current state-of-the-art. We applied CellAnimation to investigate the differences in motility between LNCaP-34 and LNCaP-17 prostate cancer cell lines, selected for difference in the levels of expression of hepsin, a type II transmembrane serine protease. Hepsin is overexpressed in over 90% of prostate cancers and correlates with tumor progression. Our lab has previously shown that hepsin cleaves laminin-332, an important protein component of the basement membrane that curbs cancer invasion and progression. Automated cell tracking and data analysis demonstrated that hepsin overexpression promotes increased cell speed and displacement but path tortuosity stays the same; net speed increase was accompanied by a switch in integrin use and a more mesenchymal morphology.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-06042012-213228 |
| Date | 05 June 2012 |
| Creators | Georgescu, Walter |
| Contributors | John P. Wikswo, Vito Quaranta, Jeffrey M. Davidson, Frederick Haselton, Shane Hutson, Adam Anderson |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-06042012-213228/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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