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As a consequence of poor perfusion and elevated acid production, the extracellular pH (pH^ex) of tumors is generally acidic. Despite this, most in vitro experiments are still performed at the relatively alkaline pH^ex of 7.4. This is significant, because slight changes in pH^ex can have profound effects on cell phenotype. In this study we examined the effects of mildly acidic conditions on the in vitro invasive potential of two human melanoma cell lines: the highly invasive C8161, and poorly invasive A375P. We observed that culturing of either cell line at acidic pH (6.8) caused dramatic increases in both migration and invasion, as measured with the Membrane Invasion Culture System (MICS). This was not due to a direct effect of pH on the invasive machinery, since cells cultured at normal pH (7.4) and tested at acidic pH did not exhibit increased invasive potential. Similarly, cells cultured at acidic pH were more …