An Assay for Analyzing Test β-Glucan as an Immunotherapy Strategy against Brain Cancer Cells
Published: May 31, 2024
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. Prepare Dulbecco's modified Eagle's medium (DM…
Declarações
The authors have nothing to disclose.
Materials
| 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 |
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An Assay for Analyzing Test β-Glucan as an Immunotherapy Strategy against Brain Cancer Cells. J. Vis. Exp. (Pending Publication), e22276, doi: (2024).