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Begin by adding a cold suspension of glioblastoma and microglial cells, each stained with a distinct fluorescent dye, into a cold polymer matrix solution.
The cold condition prevents the premature polymerization of the matrix.
Next, transfer the mixture into a porous chamber insert within a culture plate.
Incubate to allow matrix polymerization, forming a gel that embeds the cells.
Add serum-free media to the insert and serum-containing growth media to the well below, then incubate.
The chemical factors in the serum promote cell migration toward the serum-rich media.
Additionally, microglia secrete signaling molecules that interact with glioblastoma cells.
This enhances the glioblastoma cell migration to the underside of the porous insert, resembling their migration from a tumor into the surrounding tissue.
Remove the cells from the top of the chamber.
Now, fix the remaining cells with a fixative and wash them with buffer. The cells' distinct fluorescence helps track cell migration.
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