i am trying to prepare phospholipid/BSA solution to add it to cell culture. so please does anyone has experience in dissolving phospholipids in BSA but I don't want to form micelles.
Thanks in advance,
Dear Samaa,
BSA has several hydrophobic 'patches' that can bind and transport hydrophobic compounds such as lipids in the blood. From the top of my head there are six of these binding sites per albumin, having different affinity. So make sure you have done the math to ensure there is enough albumin.
Whether or not micelles will be formed is not trivial. Most phospholipids will not form micelles but vesicles, as they self-organize in bilayers. This depends mainly on the phospholipid headgroup and whether you have one or two acyl chains present. I suppose you should expect a dynamic equilibrium between lipids in vesicles and lipids bound to BSA. Only at very low [PL] is expect all PL to be BSA bound. Hope this helps.
To prevent the strong interaction between the glass surface and cultured cell or lipid membrane, the inside of the microchamber should be coated with 0.10% (w/v) BSA which is dissolved in the buffer containing approximately 0.1 M glucose or other physiological buffer you want to make.
Lipid and albumin conjugation and solubilization must be verified when preparing FA/BSA solutions. Indeed, differences in these formulations can generate disparate responses in the same system. For instance, microglial cells treated with palmitate at a FA:BSA ratio of 2:1 present an anti-inflammatory profile, while a 10:1 ratio induces a strong pro-inflammatory profile.
You can check this article. Hope this helps you. Best wishes. Article Cell culture models of fatty acid overload: Problems and solutions
@ Md Kabir, the paper you posted is really helpful and has a plenty of useful information. Many thanks.
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