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

Abstract

Source: Folgado-Dorado, C. et al., Isolation and Purification of Fungal β-Glucan as an Immunotherapy Strategy for Glioblastoma. J. Vis. Exp. (2023)

This video demonstrates an in vitro assay utilizing β-glucans to activate microglial cells for the production of superoxide radicals, a reactive oxygen species. In brain cancer cells, the generated reactive oxidants actively suppress tumor cell proliferation and induce cell death.

Protocol

In vitro study of β-glucan-induced microglia stimulation

1. Cell culture of mouse glioblastoma and microglia cells in 8-well chamber slides

NOTE: This protocol is specific for GL261 (glioblastoma) and BV2 (microglia) cell lines. However, with slight modifications, these steps could potentially be used to study other cancer and immune cell lines.

1. Prepare Dulbecco's modified Eagle's medium (DMEM) complete medium modified with L-glutamin, 4.5 g/L D-glucose, and without pyruvate. Add 10% of fetal bovine serum (FBS) and 1% penicillin/streptomycin. Pre....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
8-well chamber slides	Thermo Fisher, USA	171080
Dulbecco's modified Eagle's medium, Gluta MAXTM	Gibco, Life Technologies, Carlsbad, CA, USA	10564011
Extenda (α- Amylase/Glucoamylase)	Novozymes, Denmark
Fetal bovine serum	Gibco, Life Technologies, Carlsbad, CA, USA	A4736301
Phosphate-buffered saline	Gibco, Life Technologies, Carlsbad, CA, USA	1010-015
Penicillin/streptomycin	Sigma-Aldrich, St. Louis	P4458
Incubator	Eppedorf	Galaxy 170S