Brain Tumor Stem Cell (BTSC) Migration Assay for Drug Treatment Studies: An In Vitro Live-cell Imaging Technique to Analyze the Effect of Drug Treatment on Migration of BTSCs

Brain tumor stem cells or BTSCs, a brain tumor cell subpopulation, can initiate tumor growth and metastasis, making them promising targets for cancer therapy. To study the effect of drugs on BTSC migration in vitro, first, prepare uniform suspensions of BTSCs in a suitable media. Treat one of the suspensions with the drug of interest.

Meanwhile, obtain a multi-well chemotaxis plate consisting of membranous inserts that separate each well into upper and lower chambers. Pipette desired extracellular matrix protein to the lower chambers and membranous inserts. Incubate to facilitate the polymerization of the protein. 

Add a suitable basal media containing the desired chemoattractant to the lower chambers and place over the membranous inserts. Next, seed the drug-treated and untreated BTSC suspension into membranous inserts. Image the plate for an extended duration using live-cell microscopy.

In culture, BTSCs adhere to the protein matrix of the insert. In response to the chemoattractant concentration gradient established between the upper and lower chambers, the untreated BTSCs migrate across the protein matrix through the membrane insert pores to the lower chamber. 

However, the drug treatment negatively affects the cells' functional activity, decreasing BTSC migration. Now, using suitable software, analyze and quantify the migrated BTSCs in the lower chambers. The untreated BTSCs display increased migration, while the drug-treated BTSCs show reduced migration.