Hi, I've read numerous studies that treat SHSY5Y cells with 100-1000 uM FeCl2. In these studies, there is no mention of how this solution was prepared. I assumed they simply added iron chloride to cell media (DMEM/F12 + 10% FBS), but I'm getting tons of precipitate in my cell culture when I do this.
What am I missing? How do I treat my cells with 100 uM FeCl2? Sounds easy???
Normally, you make a stock solution in medium, which you dilute appropriately with neat medium. So, for example, if you want to treat cells with 200 µL of 0, 100, 200,...,1000 µM, you'd prepare a 1000 µM stock and add 0, 20, 40,..., 200 µL of the stock to 200, 180,..., 0 µL neat medium.
The condition for this is that the compound needs to be soluble at the maximum concentration you want to use. If it is not, you may be able to increase solubility with complex formers like EDTA or citrate. Then the concentration of this additive needs to be the same in all experiments, i.e., you add that complex former both to the Fe solution and to the neat medium at the same concentration. You'd also do a zero Fe, zero complex former control to account for the effects of the complex former alone.
Note that FeII is easily oxydised to FeIII (in particular under alkaline conditions), which is even less soluble and has different toxicological properties. So you may have to exclude all oxidants, including air oxygen, from the experiment.
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