Brain Tumor Stem Cell (BTSC) Migration Assay for Drug Treatment Studies: An In Vitro Live-cell Imaging Technique to Analyze the Effect of Drug Treatment on Migration of BTSCs
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)
In this video, we perform brain tumor stem cell migration assay to analyze the effect of the drug of interest on the migration of live brain tumor stem cells using chemotaxis migration plate. Live-cell imaging facilitates the visualization and quantification of cell migration during the assay.
Protocol
1. Brain Tumor Stem Cell Migration Assay Seed 200,000 cells in a T25 flask in 8 mL of complete media 1–2 weeks before planning to perform a migration assay. NOTE: The time between plating and performing the assay will vary depending on the growth rate of the BTSC culture used. To test the effect of a drug treatment on migration, pretreat the cells with the appropriate vehicle, such as DMSO, and drug of interest by adding applicable volumes of the vehi…
Representative Results
Figure 1: Live-cell imaging of a BTSC migration. (A) This is an example of a 96-well chemotaxis migration plate coated with a thin layer of type I collagen prior to performing the migration assay with BTSCs. The images show i) a plate with all three parts (lid, membrane insert, and reservoir) placed together, ii) all three parts separated, iii) a close-up image o…
Divulgaciones
The authors have nothing to disclose.
Materials
| Matrigel matrix | Corning | 356230 | |
| 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 | |
| Phosphate buffered saline (PBS) | Sigma | D8537 | |
| Culture grade water | Lonza BioWhittaker | 17-724Q | |
| Fetal bovine serum (FBS) | Invitrogen | 12483-020 | |
| Rat Collagen I | Trevigen | 3440-100-01 | |
| Accumax cell detachment solution | Innovative Cell Technologies | AM105 |
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Brain Tumor Stem Cell (BTSC) Migration Assay for Drug Treatment Studies: An In Vitro Live-cell Imaging Technique to Analyze the Effect of Drug Treatment on Migration of BTSCs. J. Vis. Exp. (Pending Publication), e20678, doi: (2023).