This study investigates the effects of β-glucans on microglia and their potential anti-tumoral properties against brain cancer cells. By activating microglia with β-glucans, researchers aim to understand the resulting bioactive molecules and their impact on cancer cell proliferation and apoptosis.
Take a multi-well chamber slide containing microglia monolayers.
In the test well, coat the cells with β-glucan. The control well lacks β-glucan. Incubate.
In the test well, β-glucans, pathogen-associated molecular patterns, bind to specific receptors on microglia, causing β-glucan phagocytosis.
This activates the microglia, producing and releasing bioactive molecules such as reactive oxidants and cytokines into the media.
Collect the media containing bioactive molecules from the wells and filter them to remove debris and β-glucan.
Freeze the media at low temperatures to preserve the biological activity of the bioactive molecules.
Next, thaw and add the media to a multi-well chamber slide containing monolayers of brain cancer cells. Incubate.
The bioactive molecules trigger cell signaling pathways, reducing cancer cell proliferation and inducing apoptosis.
Reduced cancer cell proliferation and increased apoptosis suggest β-glucans as agents promoting anti-tumoral properties of microglia 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.
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