Can cell migration/invasion in the presence and absence of pharmacological inhibitors be tested for 1 week using this assay?
The duration of experiment may differ depending on the cancer cell line you use, also the method and conditions (normoxia/hypoxia).
There are some different assays to test cell invasion/migration. Some of them may be done in 5-7 days (inverse invasion assay), some in 8-10 days (3D invadopodia formation), etc.
Dear Es,
The duration of invasion assays depends on the migratory abilities of your cell type of interest (there are remarkable difference among different normal and cancer cell types).
However, they typically last only a few days (2-5 days) to avoid medium replacement (not so straightforward for inverted invasion assays) and possible confounding effects due to cell proliferation (especially for spheroid assays).
Moreover, you should try to employ an extracellular matrix (e.g. collagen, fibronectin, etc.) that mimics that of your cells of interest. Consider that migration is strongly influenced by the ECM.
For some examples and further reading:
Article Initiation of lamellipodia and ruffles involves cooperation ...
Article Rapid Remodeling of Invadosomes by Gi-coupled Receptors: dis...
Article Invadosomes - shaping actin networks to follow mechanical cues
Article New insights into the formation and the function of lamellip...
Best,
Metello
To assess the migration behaviour of your cells you could start with a simple chemotaxis assay. Those are filter inserts that got in a multi well plate [i.e. 12]. You seed starved cells on top of the filter and place a typical stimulant in the bottom wells [high FBS media for example]. After you have established the number of cells that typically migrate you can specifically inhibit that migration. That should give you an idea how your cells migrate before you attempt more sophisticated live cell microscopy assays or 3D models.
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