Comparing qualitative and quantitative spectroscopic techniques for the detection of the effect of direct iron loading of mammalian cell cultures.

摘要: Iron overload augments diseases of the liver and microorganism infection as well deregulates immune system. In vitro analysis effects iron loading its chelation involves determining amount constituting overload, which metal sources cell lines to use reliable assay methods. The uptake 500 µM FeSO4 or FeEDTA by CEMss, U937 leukocytes was confirmed inductively coupled plasma-atomic emission spectroscopy (ICP-AES). Excess increased CEMss viability (assessed MTT, XTT, Trypan-and Alamar Blue) an average 18% (P = 0.034). Flow cytometry indicated dye-viable cells be undergoing apotosis/necrosis while still confirming increase (9%, P < 0.001) in excess iron-induced viability. chelator desferioxamine (DFO) when added addition Fe, reversed (and vice versa) had detrimental used on own (33% inhibition measured dyes 10.85%; 0.0427 assessed flow cytometry). 4 demonstrated different levels sensitivity detecting influence DFO but gave a related, qualitative picture ICP-AES data more quantitative.