Luciferase-tagged Glioma Stem Cell Model: A Lentivirus-mediated Method to Prepare Luciferase-tagged Glioma Stem Cells for In Vitro Therapeutic Studies
Published: April 30, 2023
Abstract
Source: Hu, Z. et al. A Rapid Screening Workflow to Identify Potential Combination Therapy for GBM using Patient-Derived Glioma Stem Cells. J. Vis. Exp. (2021)
This video demonstrates the establishment of luciferase-tagged glioma stem cells via lentivirus-mediated gene delivery. Luciferase acts as a reporter gene and helps facilitate rapid drug screening experiments.
Protocol
1. Preparing Luciferase-tagged GSCs Begin with the GSCs culture and centrifuge at 70 x g for 3 min at room temperature. Remove the supernatant and digest the cells with accutase for 4 min at 37 °C. Use a 200 µL tip and pipette repeatedly to dissociate and resuspend the cell pellet. Dilute the cells to 2 x 105 cells per well in a 12-well culture plate and culture the cells overnight. Add 30 µL luciferase-EGFP virus…
Offenlegungen
The authors have nothing to disclose.
Materials
| B-27 | Gibco | 17504-044 | 50X |
| EGF | Gibco | PHG0313 | 20 ng/ml |
| FGF | Gibco | PHG0263 | 20 ng/ml |
| Gluta Max | Gibco | 35050061 | 100X |
| Neurobasal | Gibco | 21103049 | 1X |
| Penicillin-Streptomycin | HyClone | SV30010 | P: 10,000 units/ml S: 10,000 ug/ml |
| Sodium Pyruvate | Gibco | 2088876 | 100 mM |
Tags
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Luciferase-tagged Glioma Stem Cell Model: A Lentivirus-mediated Method to Prepare Luciferase-tagged Glioma Stem Cells for In Vitro Therapeutic Studies. J. Vis. Exp. (Pending Publication), e20613, doi: (2023).