Preparation of Collagen-Coated Compartmented Culture Dishes for Neuronal Cell Culture
Transkript
Start by adding a collagen solution to a culture dish.
The solution's acidic pH prevents collagen matrix formation.
Remove the excess solution.
Place the dish in a tray along with a gauze pad soaked in ammonium hydroxide. Place a lid to contain ammonia fumes.
These vapors raise the pH, neutralizing the acidic environment.
Neutralization disrupts the electrostatic repulsion between collagen molecules, allowing them to form a stable matrix. Create horizontal scratches across the center.
Apply a non-reactive grease to the base of a compartmented chamber.
Now, attach the scratched surface of the dish to the chamber.
Flip and apply the grease to seal the central compartment.
Add media to check for leakage. Seal the leakage with grease.
This forms a three-compartment system: the central compartment for neuronal cells and the side compartments for tumors.
Neuronal cells grow along scratches to side compartments and produce a myelin sheath on axons, facilitating the study of the tumor-cell interaction.
