I was planning to simulate hypoxia chemically in cancer cells using dimethyloxalylglycine (DMOG), desferrioxamine (DFO), ciclopirox olamine (CPX) or cobalt chloride (CoCl2), due to lack of a hypoxia chamber. However, I was also wondering if the results obtained from this form of hypoxia induction comparable to hypoxic simulation in a chamber, but I only found one article for this proof related to mesenchymal stem cells: Article Hypoxia-mimetic agents inhibit proliferation and alter the m...
I have yet to find an article noting a difference between the two in cancer cells. Has anyone tried both cases and noticed a significant difference, or are they comparable? Similarly, does the agent type also affects results? I can't find any article comparing the effect of each agent in cancer cells as well. Thanks!
There are some discussions about differences between chemical hypoxia and hypoxia induced in chamber here: https://www.researchgate.net/post/Hypoxia
I hope it will be useful for you.
I'd agree with the authors above, and add; With reference to cobalt compared to hypoxia. Both are quite different, and operate via completely different mechanisms. Cobalt chloride blocks the PHD enzymes and HIF1 alpha degradation via vHL, leading to HIF1 alpha accumulation. In stark contrast, hypoxia is a lowering of one of the substrates (oxygen) required by those same PHD enzymes. A true hypoxic response is dynamic and varies with the degree (% oxygen) and duration of hypoxia - even within a constant hypoxic environment - in a manner not seen with cobalt. Both methods can elevate HIF1 alpha, but the choice really depends upon what you want to achieve with your experiments, and what resources you have available.
It's also important to keep in mind that chemical hypoxia (cobalt, DMOG..), inhibits "specifically" PHD and promotes HIFa stabilization. However, real hypoxia induces several changes independently of HIF (for example, ER stress and UPR response, diminution of ATP, AMPK activation and inhibition of mTOR, and ROS production changes). Thus, it's necessary to distinguish the role of hypoxia and the role of HIFa stabilization.
I agree with all researchers above. Briefly, in my experience i used DMOG for HIF1a studies and I used hypoxia chamber 1% O2 for the same purpose. The results showed that DMOG increased the expression HIF1a, while the chamber didn't.
I hope that is useful.
Hi! Looking at the consensus, I decided to not go through with the chemical test at all. Thanks for everyone's extra inputs, they did clear the doubt I have about their differences!
All of the advice on chemical induced hypoxia above is good, but if you don't want to add chemicals to induce hypoxia, just do it the natural way. Get a Petaka G3 cell culture cassette and follow the hypoxia protocol. You can induce to as low of levels as you want, for as long as you want, and monitor pO2 in real-time so you can correlate the actual pO2 in your media with your experimental results. Plus, the Petaka stays hypoxic for a long time outside of incubators and glove boxes so your working time is excellent.
www.petaka.com
https://www.youtube.com/watch?v=JmYj4ogtfUo
https://celartia.com/product-category/oxygen-measurements/
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