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Method Article
Claire Lugassy
University of California, Los Angeles
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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
Keywords
Extra-vascular migratory metastasis (EVMM), angiotropism, pericytic mimicry; melanoma; cancer, metastasis in vitro 3D co-culture model; real time monitoring
Figure 1
Figure 2
Authors declare no conflict of interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.mp4
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.
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Associated Publications
Human Melanoma Cell Migration Along Capillary-Like Structures In Vitro: A New Dynamic Model for Studying Extravascular Migratory Metastasis
by Claire Lugassy, Hynda K. Kleinman, Patricia M. Fernandez, +5
Journal Of Investigative Dermatology (21 November, 2013)
Could pericytic mimicry represent another type of melanoma cell plasticity with embryonic properties?
by Claire Lugassy, Bruno Péault, Madhuri Wadehra, +2
Pigment Cell & Melanoma Research (21 November, 2013)
Pilot Study on “Pericytic Mimicry” and Potential Embryonic/Stem Cell Properties of Angiotropic Melanoma Cells Interacting with the Abluminal Vascular Surface
by Claire Lugassy, Madhuri Wadehra, Xinmin Li, +8
Cancer Microenvironment (21 November, 2013)
Glioblastoma Stem Cells Generate Vascular Pericytes to Support Vessel Function and Tumor Growth
by Lin Cheng, Zhi Huang, Wenchao Zhou, +11
Cell (21 November, 2013)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Claire Lugassy
University of California, Los Angeles
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 22 Nov, 2013
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
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
Authors declare no conflict of interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.mp4
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.
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Associated Publications
Human Melanoma Cell Migration Along Capillary-Like Structures In Vitro: A New Dynamic Model for Studying Extravascular Migratory Metastasis
by Claire Lugassy, Hynda K. Kleinman, Patricia M. Fernandez, +5
Journal Of Investigative Dermatology (21 November, 2013)
Could pericytic mimicry represent another type of melanoma cell plasticity with embryonic properties?
by Claire Lugassy, Bruno Péault, Madhuri Wadehra, +2
Pigment Cell & Melanoma Research (21 November, 2013)
Pilot Study on “Pericytic Mimicry” and Potential Embryonic/Stem Cell Properties of Angiotropic Melanoma Cells Interacting with the Abluminal Vascular Surface
by Claire Lugassy, Madhuri Wadehra, Xinmin Li, +8
Cancer Microenvironment (21 November, 2013)
Glioblastoma Stem Cells Generate Vascular Pericytes to Support Vessel Function and Tumor Growth
by Lin Cheng, Zhi Huang, Wenchao Zhou, +11
Cell (21 November, 2013)