fatty acid free BSA-cell culture - (Jan/23/2012 )
I need to treat my cells Huh7 with oliec acid to induce fatty liver model. I've read in articles about fatty acid free bovine serum albumin and how they use a combination of fatty acid and FAF-BSA. I am wondering what's the point of using FAF-BSA . Do u think using FAF-BSA will minimize the amount of oliec acid the cells will expose to (which somthing we must avoid it)? There must be a reason for using it....any help in details plzzzzz
It would be great if u could recommend some refrences...
Thanks in advance
It would be so that you can control the levels of oleic acid - that way you can determine if the effect is dependent on the amount of oleic acid present and/or if the effect is due to oleic acid alone.
Thank u for ur replay but can u explain more
Presumably you are going to add oleic acid to your cells to induce the fatty liver model, as you mentioned in your first post? Oleic acid is a fatty acid - so if you use ordinary BSA, how do you know how much oleic acid it has? If you use FAF-BSA, you can add a defined amount of oleic acid, and thereby determine that the effect you are seeing is due only to the oleic acid you added, not some already present in the BSA or from some other component in the BSA.
BSA is used as a carrier of long- chain
I have also been treating cells with different types of FA (such as OA) and for this I dont use BSA. Like you, I see that people tend to use a combination of FAF-BSA however I could not find an explanation for why they do it like that. I have seen people saying that is a vehicle for lipids because it binds to the FA not allowing it to stick everywhere, well this makes sense if you adding it to living beings but if you using for cell cultures I dont see why it should matter and I will explain why:
1- If you dissolve your FA in for example Ethanol you can obtain an homogenous solution if you don't go above the solubility limit. This way you make sure you adding the amounts you want. You just need to be careful to the amount of Ethanol you adding to the cultures (ex. for COS cells you should not go above 1% (v/v)). Of course high FA concentrations are toxic to the cells but you would need to go very high since cells have mechanisms to protect against lipotoxicity (ex: storing lipids in LDs). Thid also depends in the type of cells you using.
2- If you use BSA and it binds to FA how much FA is going to diffuse to the cells? Also, if you have a BSA-FA complex this is probably less permeable to the cell membrane than the FA. In addition, cell membranes have FA transports.
In terms of references I cant help you there.