I am using Oleic (liquid) and Palmitic (powder form) acids to observe lipid droplets in HCC cell lines. Currently, I am doing research to find the best protocol to dissolve them. So far , we have tried solving the fatty acids in 20% Ethanol (heat up to 70C), then conjugate with pre-warmed (37 C)1% BSA. Make stock solutions of 250mM (With ratio of 2:1 oleic:palmitic acid)
Unfortunately, we do not see any changes in lipid accumulation. It seems the solutions hasn't been taken up by the cells.
I would like to kindly ask you all if you could provide me with a protocol or some references for handling fatty acids for cell culture?
Hi,
Could try the following:
1. Dry oleic acid under N2 gas and then reconstitute it in DMSO.
2. Dissolve palmitic acid in DMSO.
3. Add same volume (ul) of DMSO to the control.
Because fatty acid can diffuse and/or rapidly flip-flop, try adding to the cells directly. It might be taken up by the cell, but please also consider cell's toxicity by limiting the dose.
Deepti
Dry the oleic or palmitic acid and reconstitute in a solution of 0.25 mg/mL fatty acid free serum albumin made up in your Hanks Balanced Salt Solution or whatever buffer you are using with the cells.
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