I used human bone marrow aspirate to culture MSC, sometimes the sample is mixed with EDTA. I have notice these samples show different proliferation rate and morphology from the the samples without EDTA. I wonder if you have experience with this condition? Thank you!
some membrane and non membrane proteins require divalent metal cations for proper function, addition of EDTA will remove these divalent metal cations, resulting in altered protein function, which could lead to altered cell behaviour.
Hello
The addition of EDTA usually accelerates the dissociation phase through the removal the links between cells , As " Steven R Pirie-Shepherd " said remove divalent metal cations affect protein function and this affects the cells on the culture .
Good Luck
Mg2+ and Ca2+, which EDTA chelates, is required for cell adhesion, hence your 'different' morphology. This explains what EDTA is doing in your trypsin and why your PBS probably specifies that it is without calcium and magnesium. If your EDTA concentration is high enough, it will affect your culture, as you have noticed. If you are unable to avoid adding EDTA in the first place, or cannot pellet your cells to remove it once added, you could work out the final concentration of EDTA in your medium and supplement it with an equal concentration of Mg2+ and Ca2+. For example, if you work out that your EDTA final concentration is 1mM, adjust your medium so that you have an additional 0.5mM MgCl2 and 0.5mM CaCl2.
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