The chemoinvasion assay: a method to assess tumor and endothelial cell invasion and its modulation.
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.
Albini
Adriana
A
Polo Scientifico e Tecnologico, IRCCS Multimedica, Milano, Italy. adriana.albini@multimedica.it
Benelli
Roberto
R
D001485:Basement Membrane / Q000502:physiology; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002633:Chemotaxis / Q000502:physiology; D003094:Collagen / Q000494:pharmacology / Q000502:physiology; D003584:Cytological Techniques / Q000379:methods; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology*; D005109:Extracellular Matrix / Q000502:physiology; D007797:Laminin / Q000494:pharmacology / Q000502:physiology; D009361:Neoplasm Invasiveness; D009389:Neovascularization, Pathologic; D011509:Proteoglycans / Q000494:pharmacology / Q000502:physiology
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen