For a current study I would like to treat the HuH7 cell line with palmitate-BSA. I have previously synthesized palmitate-BSA via the protocol of Seahorse Bioscience (see attachment). I am wondering, however, if there are methods to improve this synthesis route or are there improved protocols for the synthesis of palmitate-BSA? This in order to optimize the uptake of palmitate-BSA by the HuH7 cells.
Dear Daan, I have also used this protocol from Seahorse, but I haven't made any improvements. You may try palmitate ethyl ester instead of palmitate-BSA. Ethyl esters of long chain fatty acids are more cell permeable than conjugates with proteins.
Dear Maja, thank you very much for your reply. Do you have any ethyl esters in mind to conjungate the palmitate with? Or do you have a protocol to prepare these palmitate ethyl esters?
Dear Daan, the best thing is that you can buy them ready to use. For example I used various fatty acid derivatives from Cayman Chemical, such as eicosapentaenoic (EPA) ethyl ester (https://www.caymanchem.com/product/10008884), DHA ethyl ester (https://www.caymanchem.com/product/9090310) or palmitate ethyl ester (https://www.caymanchem.com/product/10008202). I think Sigma Aldrich have such reagents, too. You can dissolve them in DMSO, ethanol or dimethylformamide and add these solution to the cell culture medium.I imagine that if the medium contains serum some proportion of these fatty acids can form conjugate with serum proteins. By the way, serum itself also contains fatty acids. If you do not want let the interfere with the experiment, you can use delipidized serum.
Good luck with your studies:)
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