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…
Declarações
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
Low Frequency Ultrasound Application: A Method to Determine the Effect of Sonication on Human Leukemia Cells. J. Vis. Exp. (Pending Publication), e20256, doi: (2023).