Assessing Interneuron Response to Chemoattractive Cues from Periventricular Endothelial Cells

Fix a three-compartment culture insert in a polymer-coated dish for cell attachment.

Invert the dish, mark the middle compartment, and reorient it. Add a medium containing interneurons to this compartment.

Add a medium containing human periventricular endothelial cells, or PVECs, brain vessel cells, to one of the outer compartments.

In another outer compartment, add a medium containing non-periventricular endothelial cells as a control.

Add a co-culture medium to the dish to prevent polymer drying. Incubate for cell adherence.

Disassemble the insert, creating rectangular patches of cells with small cell-free gaps between each cell type.

Refresh with the co-culture medium and incubate.

Endothelial cells trigger interneurons to migrate into cell-free spaces.

Meanwhile, PVECs release chemical attractants that enhance the migration of interneurons toward the PVEC-containing patch.

Higher interneuron migration toward the PVEC patch than the control cell patch confirms the interneuron response to chemoattractive cues from the PVECs.