H9C2 chemical-induced hypoxia conditions - (Mar/26/2008 )

Hi all,

I've recently started working on H9C2, a cardiomyoblast cell-line. I'm inducing hypoxia using hypoxia-mimetic chemicals : cobalt chloride and desferrioxamine (DFX).
From the literature, I see that most use 100uM CoCl2 and 130uM DFX. I can get results from CoCl2 (18hr incubation) when i conduct cell viability MTT assay. However, despite trying for 4-5times, I'm unable to get results from DFX. I can't figure out why? Can anyone please help?

Also, I'm still optimizing the hypoxia duration for both chemicals. Has anyone done this before? Would really appreciate any suggestions!

Thanks for your help!

Did your drugs induce HIF-1 alpha?

Hi,

Havent run my Westerns yet, will be doing it this coming week. But, from the PCR results, I was able to get HIF-1a bands; both with and without inducing hypoxia.