Co-Culturing Glioblastoma Cells on Patterned Neurons: A Technique to Monitor the Migration of Glioblastoma Cells on Rat Hippocampal Neurons In Vitro
Transcription
Glioblastoma possesses stem-like cells that migrate along the axonal tracts of myelinated neurons to invade distant locations in the brain. To recapitulate the migration of glioblastoma or GBM cells on neurons in vitro, begin by taking a glass coverslip.
Activate the coverslip by oxygen plasma treatment and prime it with a functionalizing agent. This treatment supports easy access to the chemicals in subsequent steps. Add PEGylated solution to the coverslip and incubate. This solution coats the coverslip preventing neuron adhesion at random places.
Dispense a photoinitiator polymer onto the coverslip and mount it in the imaging chamber. Place the chamber on the stage of a fluorescence microscope. Once inside, the UV illumination activates the photoinitiator molecules locally at the designated areas corresponding to the envisioned micropatterns.
The activated photoinitiator molecules cleave the underlying PEG layer, generating the desired pattern. Now, add the laminin solution and incubate. Laminin – a glycoprotein – gets deposited at specific places where the PEG layer has been removed. Seed the media containing rat hippocampal neurons onto the coverslip.
Incubate to allow adherence of the neurons onto the laminin-coated arrays to obtain neuron-micropatterned coverslips. Deposit a suspension of fluorescently-labeled GBM cells over the micropatterned neurons. Image to see the fluorescent GBM cells migrating on neurons.
