serum free culture - doubts about adaptation n antibiotics (Jun/11/2007 )

Hello All,

I am going to use a serum free media for the culture of monocytes and macrophages.
https://catalog.invitrogen.com/index.cfm?fu...Description=328

It doesnt contain antibiotics and I suppose there are some issues with the use of antibiotics in serum free media.
we use 1% penstrep with the serum media. what % do i use in serum free media?
Does anyone have any suggestion and tips regarding culture in serum free media?

Also, I ll be taking these cells from serum media to serum free media.
Can I take them directly or is it necessary to follow the adaptation protocol?
Can I mix the serum and serum free media?

Thanks !

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No problem, use the same % of your antibiotic mixture for serum free media

Concerning culturing on serum free media you should be carefull and adopt your culture for serum free media. Of course it depends on your culture first of all. In general If you've used 10% FBS so take 5% for 2-3 days, then analyse your cells behavior, If good enough decrease till 2% for a week and then decrease till 0% but add in media BSA till 0.1% when culturing on serum free media

Good luck!

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We use serum free media for neuronal cultures. We add the same 1% pen/strep in the serum free media.

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how about antimycotics?
Can we add Fungizone also?

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We dont add antimycotics. But I dont see why that should be a problem.

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BIG POTENTIAL PROBLEM

Fungizone (Amphotericin is a very dirty compound and can inhibit many secondary messaging systems. We had a particular problem with the inhibition of Nitric oxide release from endothelial cells.

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Then what should be done to avoid fungus growth?
(other than the practice of good aseptic technique)

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