Low Frequency Ultrasound Application: A Method to Determine the Effect of Sonication on Human Leukemia Cells
Published: April 30, 2023
Abstract
Source: Trendowski, M. et al. Generation and Quantitative Analysis of Pulsed Low Frequency Ultrasound to Determine the Sonic Sensitivity of Untreated and Treated Neoplastic Cells. J. Vis. Exp. (2015).
This video describes the selective damage of human leukemia cells by using low frequency ultrasound. This application of low frequency ultrasound in a preclinical in vitro condition demonstrates that cell destruction can be attained in order to examine and compare the effects of sonication on leukemia and normal cells.
Protocol
1. Preparation of Cells and Medium Prepare the medium by using 240 ml Iscove’s Modified Dubecco’s Medium (IMDM) and 50 ml fetal bovine serum. Do not thaw fetal bovine serum in a hot water bath, as rapid increases in temperature can lead to a compromise in serum stability. Combine 240 µl gentamicin 50 mg/ml and 5 ml penicillin/ streptomycin 100x in a standard culture flask, and add to the medium. Store the medium at 4°C. Seed U937 cells at ~4 x 104 ce…
Divulgations
The authors have nothing to disclose.
Materials
| Iscove's Modified Dulbecco's Medium w/ NaHCO3 & 25mM Hepes | Life Technologies | 12440079 | |
| Fetal Bovine Serum | Sigma-Aldrich | 12105 | |
| Gentamicin 50 mg/ml | Sigma-Aldrich | G1397 | |
| Penicillin/Streptomycin 100x Solution | Life Technologies | 10378-016 | |
| Bio-Rad TC20 Automated Cell Counter | Bio-Rad | 145-0102 | |
| Trypan Blue Solution | Sigma-Aldrich | T8154 | |
| Falcon 50 ml & 25 ml Vented Culture Flasks | Fisher Scientific | 353082 | |
| 100 μl Micropipette | Wheaton | 851164 | |
| 1,000 μl Micropipette | Wheaton | 851168 | |
| U937 Human Monocytic Leukemia Cells | ATCC | CRL1593.2 | |
| THP1 Human Monocytic Leukemia Cells | ATCC | TIB-202 | |
| Branson SLPe 40kHz Cell Disruptor with 3" (25mm) Cuphorn | Branson Ultrasonics | 101-063-726 | sonication device |
| Beckman-Coulter Z2 Cell Sizer with AccuComp® Software | Beckman-Coulter | 6605700 | |
| Seal-Rite 1.5 ml Microcentrifuge Tubes | USA Scientific | 1615-5510 | |
| Beckman-Coulter Accuvette ST 25 ml Vials and caps | Beckman-Coulter | A35473 | |
| AccuJet Pro Auto Pipet | BrandTech Scientific | 26330 | |
| USA Scientific 10 ml Disposable Serological Pipets | USA Scientific | 1071-0810 | |
| Tip One 100 μl and 1,000 μl Filter Tips | USA Scientific | 1120-1840, 1126-7810 | |
| BioRad Dual Chamber Counting Slides | Bio-Rad | 145-0015 | |
| 20 ml Glass Scintillation Vials | Sigma-Aldrich | Z190527 | |
| Wheaton 250 ml Flasks | Sigma-Aldrich | Z364827 |
Citer Cet Article
Low Frequency Ultrasound Application: A Method to Determine the Effect of Sonication on Human Leukemia Cells. J. Vis. Exp. (Pending Publication), e20256, doi: (2023).