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).

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