Matrix-Based Tumor Invasion Model Using Microglia Glioblastoma Co-Culture

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.

Thaw the matrix mix at four degrees Celsius overnight. Next, prepare a 10 mg/ml concentration of matrix in the appropriate media on ice. Then, harvest the labeled cells for suspension in matrix as previ