Monitoring extra-vascular migratory metastasis (EVMM) of migrating cancer cells using an in vitro co-culture system
The mode in which cancer cells migrate away from the primary tumor is not fully understood. In conjunction with intra-vascular cellular migration, we recently proposed extra-vascular migratory metastasis (EVMM) as a means for cancer cells to venture away from the primary tumor via crawling along the abluminal vascular surface. Here, we propose a protocol that builds on a previous in vitro angiogenesis assay. This protocol details a co-culture approach to monitor fluorescence-tagged migrating cancer cells in the presence of vascular structures. This fluorescence based single cell co-culture approach to monitor cancer cell migratory behavior in real time provides a quantitative approach to decipher the cell decision making processes that cancer cells engage in when undergoing phenotypic switching and exhibiting metastasis like dynamics. This assay can also be adapted for high-throughput analysis and for therapeutic screening of cancer cell metastasis targets
Figure 1
Figure 2
This is a list of supplementary files associated with this preprint. Click to download.
Video 1 Migration of melanoma cancer cells on surface of endothelial tubules corresponding <iframe width="480" height="360" src="//www.youtube.com/embed/WxiIvpxAekQ" frameborder="0" allowfullscreen></iframe> Co-culture of GFP C8161 melanoma cells with endothelial tubules on BME. Time Lapse 5 hours to 9 hours (4 hours). The two GFP-labeled melanoma cells crawling along the endothelial tubule is shown. The melanoma cells migrate in the same direction, from right to left. Single cell analysis reveals elongated cellular structures and cell protrusions along the abluminal surfaces of the endothelial tubule suggesting EMT phenotypic switch.
Posted 22 Nov, 2013
Monitoring extra-vascular migratory metastasis (EVMM) of migrating cancer cells using an in vitro co-culture system
Posted 22 Nov, 2013
The mode in which cancer cells migrate away from the primary tumor is not fully understood. In conjunction with intra-vascular cellular migration, we recently proposed extra-vascular migratory metastasis (EVMM) as a means for cancer cells to venture away from the primary tumor via crawling along the abluminal vascular surface. Here, we propose a protocol that builds on a previous in vitro angiogenesis assay. This protocol details a co-culture approach to monitor fluorescence-tagged migrating cancer cells in the presence of vascular structures. This fluorescence based single cell co-culture approach to monitor cancer cell migratory behavior in real time provides a quantitative approach to decipher the cell decision making processes that cancer cells engage in when undergoing phenotypic switching and exhibiting metastasis like dynamics. This assay can also be adapted for high-throughput analysis and for therapeutic screening of cancer cell metastasis targets
Figure 1
Figure 2
© Research Square 2021