I have seen research papers using concentration much higher than 18uM and no one has reported that they encountered similar problem. I have made a 10mM stock (in DMSO) and dilute it with DMEM to 18uM for my cell assay. However, I can observe there were neelde-shaped crystals after incubating for 48h. Does anyone know what happened?
Based on the information provided, a few possibilities come to mind:
- Lenvatinib solubility - Lenvatinib is poorly soluble in aqueous solutions like DMEM. At higher concentrations, lenvatinib may precipitate out of solution and form crystals. 18 μM is approaching the edge of lenvatinib's solubility limit in DMEM. Using a lower concentration may prevent crystal formation.
- Stock solution - If the 10 mM DMSO stock was not adequately dissolved, undissolved lenvatinib could seed crystal growth when diluted into DMEM. Sonicating or vortexing the stock solution may help ensure full dissolution. Freshly making the stock may also help.
- Temperature - Cooler temperatures decrease solubility and could promote crystal formation during the 48h incubation. Maintaining the culture at 37°C throughout could help.
- Container - Certain plastics can interact with hydrophobic compounds like lenvatinib and cause precipitation. Switching to glass or polypropylene containers may prevent this.
- DMEM components - Salts or other components in DMEM may interact with lenvatinib and reduce its solubility. Trying a different aqueous media like PBS may be informative.
So in summary, 18 μM lenvatinib is likely near the solubility limit in DMEM. Careful stock solution preparation, optimal temperature, and evaluating different media/containers could help prevent crystal formation. Let me know if any of those suggestions help or if you!
You could also try making a lower concentration of lenvatinib in DMSO and then adding more volume to the DMEM to get the 18uM concentration. Most cells will tolerate a larger dose of DMSO. Up to a 1:10 dilution.
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