Abstract 2370: Single cell analysis using a microscale migration slide

Abstract

Abstract Cell migration occurs in a variety of biological processes including wound healing, embryonic development, and immune responses. Existing chemotaxis assays used to assess cell migration employ the Boyden chamber which lack real-time imaging capability and does not maintain well defined biomolecular gradients. To overcome these limitations, a cell migration assay was developed which enabled real-time monitoring of single cells on a slide-like platform that maintained a stable linear gradient. Fluorescence measurements with albumin (MW=66kDa) conjugated with FITC demonstrated that the concentration profile was linear over the 1 mm observation area. This linear concentration gradient was established after 1 hour incubation and was stable for over 24 hours. The microscale migration slide was used to assess the effects of serum concentrations (0, 2, 5, 10, and 15%) on the migration propensity of four different cell lines that were grown on collagen. The cell lines included cancer cell lines with known invasive properties (fibrosarcoma cancer cells HT1080 and breast cancer cells MDA-MB-231), along with HUVECs which are angiogenic cells involved in tumor metastasis and a control cell line, NIH 3T3 fibroblast cells. Cells were incubated in the experimental conditions overnight (12 hours) with images captured every 15 minutes. Migration of single cells was monitored and imaged in real-time and the percentage of cells that exhibited positive migration towards the chemotactic source was calculated. Interestingly, we observed that the highest percentage of migratory HUVEC cells was obtained using 2% serum whereas the other three cell lines exhibited increased migration with increasing serum concentration. Under similar culture conditions of either 10 or 15% serum, MDA-MB-231 possessed the highest migration index, followed by NIH 3T3 fibroblasts and HT1080 cells whereas HUVEC cells did not migrate at all under these high serum conditions. Extensive testing over a range of conditions with different users indicated remarkable consistency in the data generated and suggests that the migration slide is a useful tool to monitor the effects of chemoattractants on adherent cell lines. Overall the results provide a reference point from which to build upon future studies aimed at comparing the effects of signaling molecules and growth factors on the migration propensities of cancer cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2370. doi:10.1158/1538-7445.AM2011-2370