An Assay for Analyzing Test β-Glucan as an Immunotherapy Strategy against Brain Cancer Cells

Activate the microglia by coating the BV-2 microglial cells for 72 hours with 0.2 milligrams per milliliter concentration of four different beta-glucans isolated from P. ostreatus, P. djamor, G. lucidum, and H. erinaceus. After 72 hours, collect the supernatant with a pipette, and pass the remaining volume through a 0.20-micrometer syringe filter. Then, freeze the supernatant at -80 degrees Celsius for at least 24 hours. To treat GL261 with the pre-activated microglia-conditioned medium, add 250 microliters of beta-glucan-treated microglial medium to 80% confluent GL261 cells for 72 hours.