2.3. GBM Organoid Fabrication
Bioengineered hydrogels are a widely used tool in the tissue engineering
field. They serve as a biomimetic scaffold that supports cells and can
deliver a wide range of bioactive agents to the embedded cells. The
hydrogel used in these studies is composed of thiolated-hyaluronic acid
(HA), thiolated-gelatin, and polyethylene glycol diacrylate (PEGDA)
dissolved in water containing 0.1% w/v photoinitiator (Irgacure 2959)
to a final concentration of 1%w/v. The 3 components are combined in a
2:2:1 ratio (HA:Gelatin:PEGDA) by volume. Cells are then suspended
within the solution at a concentration of 5 million cells/ml. A 10 μL
droplet of hydrogel solution is pipetted onto each well of a PDMS coated
48-well resulting in organoid constructs containing 50,000 cells per
organoid. The organoids are then cured with 365nm wavelength UV light
for 2 seconds.