So I am trying to induce hypoxia in some mesenchymal stem cells using CoCl2. I am wondering if I need to change the media (containing CoCl2) after the induction/treatment time before assaying for any biomarkers or I can just use the culture media as it is (that is, still has the CoCl2 in it).
Thank you.
Dear Dr. Maryam Salaudeen
Yes, hypoxia induced by CoCl2 is reversible. CoCl2 is a commonly used hypoxia-mimetic agent that artificially induces hypoxia and can block the degradation and thus induce the accumulation of HIF-1α protein in normoxic conditions.
Cobalt exerts hypoxic action by occupying the iron-binding site on the PHDs, thus inhibiting the interaction between pVHL and hydroxylated HIF-1α. By using such a mechanism, CoCl2 artificially induces hypoxia by blocking HIF-1α degradation and favouring protein accumulation. The stabilization of HIF-1α is essential as it acts as a transcription factor regulating various genes.
If CoCl2 is removed, HIF1α will no longer be stable in normoxic conditions. So, you may just use the culture media as is (containing CoCl2) before assaying for biomarkers.
Regards,
Malcolm Nobre
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