Takara Bio, Inc.
Premixed WST-1 Cell Proliferation Reagent
630118
This reagent was used to evaluate the proliferation of two sarcoma cell lines, a parental and a clone with acquired resistance, in the presence of drug. This was done in order to establish the parental line's sensitivity to the drug as well as prove the resistance acquired by exposing the clone to increasing concentrations of the drug over time. This reagent was chosen for ease of use and consistency of results as these proliferation assays are done in high volumes in our laboratory.
Cell Proliferation
Established sarcoma cell line and its drug-resistant clone, generated through exposure to increasing concentrations of drug over time
Cells from both parental and resistant clones were plated at 3000 cells/well in a 96-well plate and allowed to attach overnight at 37'C in a 5% CO2 incubator. The next day, drug was added in dilutions from 0-15 uM and the cells were returned the incubator. After 72 hours, WST-1 was added to the culture plates and incubated for 2 more hours at 37'C. After incubation, results were read on a multimode detector per manufacturer's directions and the results graphed using Prism graphing software.
Product protocol calls for 10 uL/well which is below some automated pipettors dispensing capabilities. Mixing the WST-1 with a suitable amount of media (2:1 WST-1:Media in our case to dispense 15 uL) to achieve minimum dispensing volume works well.
Experiment performed in triplicate, results were corrected for background and normalized by percentage to each plate before combining for analysis. Results show a strong proliferative/survival advantage of the clone over the parental cell line in the presence of drug, showing acquired resistance to the drug.
None
Easy to use, consistent and reproducible.
Slightly expensive.
Takara/Clontech's premixed WST-1 reagent simplifies high throughput proliferation/survival assays while giving consistent results.