Jj. Gildea et al., Transmembrane motility assay of transiently transfected cells by fluorescent cell counting and luciferase measurement, BIOTECHNIQU, 29(1), 2000, pp. 81-86
Current in vitro assays used in assessing tumor motility could be improved
by the development of a simple technique that would facilitate studies of t
he impact of specific genes on pharmacologically altered chemotaxis. We dev
eloped a technique that improves on the classic transwell assay by using fl
uorescence and luminescence to assess chemotaxis. In this transient transfe
ction system, co-transfection of a reporter construct and a gene with an un
known impact on motility are coupled with biochemical assays to quantitate
the number of cells that have received a transferred gene, which subsequent
ly, crosses the membrane. This assay was found to be less variable than the
conventional transwell chamber and is easily adaptable to studies of cell
motility or cell invasion. We also demonstrate that this assay can detect t
he effect of both genetic and pharmacological inhibition of motility alone
and in combination. It therefore has the potential to reveal additive or sy
nergistic effects.