Hypothesis Effect of Extracellular Molding On Cancer Cell Morphology and Behavior Elizabeth L. Smith, RET Fellow 2010 West Aurora High School RET Mentor: Dr. Michael Cho, PhD NSF- RET Program HypothesisAbstract Conclusion Material and Methods Results  NSF Grant CBET-EEC  Prof. A. Linninger, RET Program Director  Dr. Michael Cho, Research Mentor  Brandon Lutz, fellow researcher  University of Illinois- Chicago Acknowledgements References 1. Britland, S., Perridge, C., Denyer, M., Morgan, H., Curtis, A., & Wilkinson, C. (March 01, 1997). Morphogenetic guidance cues can interact synergistically and hierarchically in steering nerve cell growth. Experimental Biology Online, 1, 2, Curtis, A., & Wilkinson, C. (January 01, 1997). Topographical control of cells. Biomaterials, 18, 24, Rapier, R., Huq, J., Vishnubhotla, R., Bulic, M., Perrault, C. M., Metlushko, V., Cho, M.,... Glover, S. C. (January 01, 2010). The extracellular matrix microtopography drives critical changes in cellular motility and Rho A activity in colon cancer cells. Cancer Cell International, 10. The environment around a cell exerts a large amount of influence over the cell’s shape and behavior. Studies in the past have shown that cells adapt both to the chemical and the topographical environment around them (1,2). Other studies have shown that the extracellular environment can change cell behavior such as motility (3). This experiment attempted to use extra-cellular, topographical cues to change cell morphology and behavior of four lines of cancer cells. Each cell line was fixed and used to create a PDMS mold. Invasive cells were seeded onto non-invasive molds and non- invasive cells were seeded onto invasive molds, then analyzed after 1 week. Actin filaments and focal adhesions were stained, recorded, and analyzed. RESULTS & ANALYSIS Use of processes similar to this could have many applications in bio-engineering of post-cancer prosthetics and implants by micro-texturing of surfaces to induce the change of invasive cells to non-invasive behavior. Four cell lines were used: NCM460 – Noninvasive colon cancer SW620 – Invasive colon cancer MCF7 – Noninvasive breast cancer MN231 – Invasive breast cancer Within the same tissue type: Invasive cells were grown on molds of noninvasive morphology Noninvasive cells were grown on molds of invasive morphology Analysis Florescent microscopy Visualizing actin filaments and focal adhesions Images were colored and overlaid using Metamorh Qualitative analysis of results Preliminary results show that: A select number of individual cells did grown within the molded substrate Cell acted differently on glass substrate than they did on PDMS Molds/Pattern could clearly be seen on most PDMS samples under DIC Cells grew as expected on glass substrate; each with a unique morphology Insert figure with all 4 on glass Cells grew differently on the unpatterned PDMS Insert figure with all 8 on glass & unpatterned PDMS Colon cell lines grew into large, colonial clumps; few individuals Insert figure with unpatterned PDMS & patterned Breast cell lines did not grow at the expected rate; high cell mortality Insert figure with unpatterned PDMS & patterned No apparent difference in cell preference between patterned and unpatterned. We shall see…. Invasive Cancer Cell Glass Substrate 1. Cells are grown on glass substrate 2. PDMS mold is created 3. Different cells are seeded 4. Cells are grown 1 week 5. Cells are analyzed for changes in structure & morphology PDMS Substrate Noninvasive Cancer Cell Cells will change morphology based on the substrate molded by cell lines of differing morphology