JoVE Encyclopedia of Experiments
Neuroscience
0 views • 3:01 min • April 28th, 2025
Begin with a flat-bottom multi-well plate coated with a basement membrane matrix, which contains extracellular polymer and adhesion proteins that mimic the tumor microenvironment's matrix.
Introduce a media containing a neurosphere of glioma cells derived from a pediatric brain tumor, ensuring it is centrally positioned in the well.
The matrix promotes the neurosphere attachment to the well.
Add a background suppressor reagent to minimize non-specific fluorescence.
Next, introduce green fluorophore-conjugated anti-CD44 antibodies.
Transfer the plate to a live-cell imaging system housed within an incubator that maintains the cell's physiological conditions.
Over time, glioma cells express the cell-adhesion protein CD44, which interacts with the adhesion proteins.
This activates the cell signaling pathway and facilitates cell migration from the neurosphere.
The fluorophore-conjugated anti-CD44 antibodies bind to the expressed CD44 on the glioma cells, making them fluoresce.
The appearance of green fluorescent cells moving away from the neurosphere confirms the glioma cell migration.
Once the wells are coated with BMM, cut a P200 tip. Take 50 microliters of the cell medium and NS from each sel
View the full transcript and gain access to thousands of scientific videos