Determining Optimal Inhibitory Tumor Treating Field Frequency for Cancer Cells: A Non-invasive In Vitro Method to Assess the Effect of TTFields on Cancer Cell Viability
Published: April 30, 2023
Abstract
Source: Porat, Y. et al. Determining the Optimal Inhibitory Frequency for Cancerous Cells Using Tumor Treating Fields (TTFields). J. Vis. Exp. (2017)
This video demonstrates the effect of TTField application on the growth of ovarian cancer cell lines in vitro. This assay can help determine the optimal frequency of the electric field that can lead to a maximum reduction in the number of cancer cells.
Protocol
1. TTFields In Vitro Application System - Base Plate and Dish Maintenance Rinse dishes and dish covers under running tap water. Then, rinse with deionized water and air-dry face down. If a chemotherapeutic agent/drug has been added to dishes in a prior experiment, fill each dish with a 5% light detergent solution and leave overnight. Rinse the dishes thoroughly under running tap water to avoid any traces of detergent inside the dish. Rinse the dishes and di…
開示
The authors have nothing to disclose.
Materials
| Invitro system and software | Novocure | ITG1000 and IBP1000 | Each unit contains 1 TTFields generator, 1 base plate, 8 TTFields dishes with covers and 1 flat cable. |
| Sterilization bags | Westfield medica | 24882 | |
| Plastic cover slides | Thermo Scientific (NUNC) | 174977 | Pre treated and sterilized |
| Glass cover slides | Thermo Scientific (Menzel-Gläser) | CB00220RA1 | Sterilize if necessary |
| Dulbecco’s modified Eagle’s medium | Biological Industries (Israel) | 01-055-1A | Warm in 37 °C water bath before use |
| RPMI 1640 | Gibco | 21875-034 | Warm in 37 °C water bath before use |
| Fetal Bovine Serum (FBS) | Biological Industries (Israel) | 04-007-1A | Warm in 37 °C water bath before use |
| L-Glutamine 200 mM (100x) | Gibco | 25030-029 | |
| Pen/Strep (10,000 U/mL Penicillin, 10,000 µg/mL Streptomycin) | Gibco | 15140-122 | |
| Sodium Pyruvate solution 100 mM | Biological Industries (Israel) | 03-042-1B | |
| Hepes buffer 1 M | Biological Industries (Israel) | 03-025-1B | |
| Insulin solution from bovine pancreas | Sigma-Aldrich | 10516-5ML | |
| 0.25% Trypsin/EDTA | Biological Industries (Israel) | 03-050-1B | Warm in 37 °C water bath before use |
| Methanol | Merck | 1.06009.2511 | Cool to -20 °C in the freezer before use |
| PBS | Biological Industries (Israel) | 02-023-1A | Without calcium and magnesium |
| A2780 | ECACC | 93112519 | Grow in RPMI 1640 supplemented with FBS (10%), pen/strep (100 U/ mL / 100 µg/mL), sodium pyruvate (1 mM) and Hepes buffer (12 mM). |
| F98 | ATCC | CRL-2397 | Grow in Dulbecco’s modified Eagle’s medium supplemented with FBS (10%), pen/strep (100 U/mL / 100 µg/mL), sodium pyruvate (1 mM) and glutamine (2 mM). |
| Ovcar-3 | ATCC | HTB-161 | Grow in RPMI 1640 supplemented with FBS (20%), pen/strep (100 U/ mL / 100 µg/mL), sodium pyruvate (1 mM), Hepes buffer (12 mM) and insulin (10 µg/mL). |
| U-87 MG | ATCC | HTB-14 | Grow in Dulbecco’s modified Eagle’s medium supplemented with FBS (10%), pen/strep (100 U/mL / 100 µg/mL), sodium pyruvate (1 mM) and glutamine (2 mM). |
| refrigirated CO2 incubator | CARON | 7404-10-3 | |
| Laminar flow cabinet | ADS Laminair | Bio12 and VSM12 |
