Brain Tumor Stem Cell (BTSC) Neurosphere Invasion Assay: An In Vitro Assay to Analyze the Effect of Drug Treatment on BTSC Invasion from Neurosphere
Published: April 30, 2023
Abstract
Source: Restall, I. et al. Live-Cell imaging assays to study glioblastoma brain tumor stem cell migration and invasion. J. Vis. Exp. (2018)
This video demonstrates the in vitro brain tumor stem cell (BTSC) invasion assay to determine the effect of drug treatment on the invasion capacity of BTSCs from the neurosphere via live-cell imaging technique.
Protocol
1. Brain Tumor Stem Cell Neurosphere Invasion Assay Seed 200,000 cells in 8 mL of complete media in a T25 flask 1–2 weeks before planning to perform an invasion assay. NOTE: The timing from plating to performing the assay will depend on the proliferation rate of the BTSC culture used. For pretreated cells, add the compound of interest to the flask at the desired concentration for the predetermined time course before the neurospheres are ready to be pl…
Declarações
The authors have nothing to disclose.
Materials
| Rat Collagen I | Trevigen | 3440-100-01 | |
| NaOH | VWR | BDH3247-1 | |
| Acetic Acid | Millipore Sigma | AX0073-6 | |
| 96-well plates | Eppendorf | 30730119 | |
| T25 Nunc EasYFlask | ThermoFisher | 156367 | |
| Falcon 15 mL conical tube | ThermoFisher | 352096 | |
| Incucyte ClearView 96-Well Chemotaxis plate | Essen Bioscience | 4582 | |
| IncuCyte Zoom Live-Cell Analysis System | Essen Bioscience | 4545 | |
| IncuCyte Zoom Live-Cell Analysis Software | Essen Bioscience | 2016A Rev1 | |
| Penicillin, streptomycin | Sigma | P4333 | |
| Phosphate buffered saline (PBS) | Sigma | D8537 | |
| Fetal bovine serum (FBS) | Invitrogen | 12483-020 |
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Brain Tumor Stem Cell (BTSC) Neurosphere Invasion Assay: An In Vitro Assay to Analyze the Effect of Drug Treatment on BTSC Invasion from Neurosphere. J. Vis. Exp. (Pending Publication), e20679, doi: (2023).