Mar 05, 2026
Comparison of drug sensitivity between MTSs and their parental cells
- Guangli Suo1
- 1CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences
- MTS
Protocol Citation: Guangli Suo 2026. Comparison of drug sensitivity between MTSs and their parental cells. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6wo5rgqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 05, 2026
Last Modified: March 05, 2026
Protocol Integer ID: 262572
Keywords: comparison of drug sensitivity, changes in cellular metabolic activity, drug sensitivity, cellular metabolic activity, parental cells cell viability, drug, mtss, cell, celltiter
Abstract
Cell viability was assessed using the CellTiter-Glo (CTG) assay to quantify drug-induced changes in cellular metabolic activity.
Guidelines
1. Seed 2,000 MTS derived cells and RCs in each well of 96-well plates and culture overnight.
2. Treat cells with drugs in culture medium at specified concentrations.
3. Measure cell viability after 48 hours using Celltiter-Glo.
4. Perform immunofluorescence staining on MTS-M231 and parental M231 cells.
5. Fix cells with 4% paraformaldehyde, block with 1% BSA, and incubate with primary antibody at 4°C overnight.
6. Use Hoechst 33342 dye for nuclear staining.
7. Capture images using confocal microscopy and analyze with ImageJ software.
Materials
- MDA-MB-231 (M231)
- MCF7
- MDA-MB-435S
- BT-549
- AU-565
- MDA-MB-453
- MDA-MB-435
- 5-Fluorouracil
- Cisplatin
- Carboplatin
- Epirubicin
- Palifosfamide
- Gemcitabine
- Docetaxel
- Paclitaxel
- Vinorelbine
- Trastuzumab
- Celltiter-Glo (Promega)
- γ-H2AX-Alexa488-conjugated antibody (Abcam)
- Hoechst 33342 dye
- Confocal microscopy (OLYMPUS, Japan)
- ImageJ software
Troubleshooting
Comparison of drug sensitivity between MTSs and their parental cells
Seed 2,000 MTS derived cells and RCs obtained from MWC-chips in each well of 96-well plates and culture overnight.
Treat the cells with drugs in culture medium at the following concentrations: 2.5 µM 5-Fluorouracil, 20 µM Cisplatin, 30 µM Carboplatin, 2.5 µM Epirubicin, 3.5 µM Palifosfamide, 0.1 µM Gemcitabine, 0.2 µM Docetaxel, 0.5 µM Paclitaxel, 2 nM Vinorelbine, and 20 µg/ml Trastuzumab.
After 48 hours of treatment, measure cell viability using Celltiter-Glo (Promega).
Perform immunofluorescence (IF) staining on MTS-M231 cells and their parental M231 cells, with or without cisplatin treatment, to visualize γ-H2AX as a direct biomarker of DNA double-strand break formation.
Seed cells onto glass cover slips and culture for 48 hours.
Fix cells with 4% paraformaldehyde, block with 1% BSA for 1 hour, and incubate with primary antibody (γ-H2AX-Alexa488-conjugated, Abcam) at 4°C overnight.
Use Hoechst 33342 dye for nuclear staining.
Capture images using confocal microscopy (OLYMPUS, Japan) and analyze using ImageJ software.