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The chemoinvasion assay: a method to assess tumor and endothelial cell invasion and its modulation

Abstract

Invasive and metastatic cells, as well as endothelial cells, must cross basement membranes (BMs) in order to disseminate or to form new blood vessels. The chemoinvasion assay using the reconstituted BM Matrigel in Boyden blind-well chambers is a very rapid, easy, inexpensive and flexible test that can be used to quantify the invasive potential of most cell types; it can be applied to detect the migratory activity associated with matrix degradation and can also be adapted to study the selective degrading activity on different matrix substrates. Transwell inserts can also be used. Once the optimal experimental conditions are empirically determined for specific cellular models, the chemoinvasion assay can be used for the screening of inhibitors of invasiveness and angiogenesis, or to select for invasive cellular populations. This protocol can be completed in 9 h.

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Figure 1: The use of blind wells in chemotaxis and chemoinvasion: the only difference between a chemotaxis (left) and an invasion chamber (right) is the addition of a Matrigel coating to chemotaxis filters (cyan).
Figure 2: These custom-made filter boxes are useful for singularly managing the filters for chemotaxis and chemoinvasion tests, but unfortunately are not commercially available and must be fabricated.
Figure 3: The appearance of migrated and invaded cells of different origin on filter surface.
Figure 4: An example of molecular correlates of invasive activity.

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Acknowledgements

This protocol has been supported by grants from the Compagnia di San Paolo, the AIRC (Associazione Italiana per la Ricerca sul Cancro), the MIUR Ricerca Finalizzata and ISS progetto Italia-USA.

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Correspondence to Adriana Albini.

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Albini, A., Benelli, R. The chemoinvasion assay: a method to assess tumor and endothelial cell invasion and its modulation. Nat Protoc 2, 504–511 (2007). https://doi.org/10.1038/nprot.2006.466

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