Cobalt (II) Chloride hexahydrate (CoCl2 • 6H2O, MW=237.9) is a chemical inducer of hypoxia-inducible factor-1.3 8. This product is soluble in water (100 mg/ml), yielding a clear, red solution.

Prepare a 25mM stock solution in sterile dd water, (prepare immediately before use)

Use CoCl2 at the final concentration of 100μM in your regular cell culture media to induce hypoxia.

Add the CoCl2 containing media to your cells and incubate the cultures for 24hours in a conventional incubator (37°C; 5% C02).

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3217626/

You could try cobalt chloride (CoCl2). It has been extensively used in the literature to "simulate" hypoxic conditions. In our lab we have used it with PMA-treated THP-1 cells and, indeed, it seems to work. Search for any relevant literature for your case and give it a try.

you could use Nitrogen cylinder to reduce the co2 level and induce hypoxia. You could check for HIF 1 expression to confirm.

We usually worked with a small hypoxia chamber (like this one http://www.brincubator.com/hypoxiachamber.htm). The cells were incubated in a Special medium for hypoxic condtions in order to keep a stable pH. Recently, also hypoxia bags were used, there like described in http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3902232/

As suggested earlier, CoCL2 works for simulating hypoxia. I have done some experiments and it was reproducible with an actualy hypoxia chamber.

Cobalt (II) Chloride hexahydrate (CoCl2 • 6H2O, MW=237.9) is a chemical inducer of hypoxia-inducible factor-1.3 8. This product is soluble in water (100 mg/ml), yielding a clear, red solution.

Prepare a 25mM stock solution in sterile dd water, (prepare immediately before use)

Use CoCl2 at the final concentration of 100μM in your regular cell culture media to induce hypoxia.

Add the CoCl2 containing media to your cells and incubate the cultures for 24hours in a conventional incubator (37°C; 5% C02).

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3217626/

It depends on what you want to study. CoCl2 or prolyl hydroxylase blockers () will stabilise the hypoxia-inducible factor and mimic the hypoxia-induced changes in gene expression. Inhibliors of the electron transduction chain will mimic severe hypoxia at the metabolic level. Mild oxidation and ATP depletion will induce redox-response similar to hypoxic one.. H2S/CO disbalance in favor of H2S (NaHS addition) will also trigger certain pattern resembling acute hypoxia. Sodium dithionite will remove oxygen from your culture medium acutely (e.g.  J R Soc Interface. 2015 Apr 6;12(105))... Each of the approaches has its disadvantages as they only mimic a part of hypoxic response pattern. I can give you some references when needed as soon as you decide what you want to achieve..,

Please read the following articles

•    Induction and Testing of Hypoxia in Cell Culture
•   Study of Cellular Responses under Chemically Induced Hypoxia
• Chemical Induced Hypoxia Differentially Affects Gene Expression in Human Neuronal Cells
•  Cobalt chloride, a chemical inducer of hypoxia-inducible factor-1α in U251 human glioblastoma cell line
• CoCl2, chemical inducer of Hif1-alpha, and CD44 expression in salivary glands malignant neoplasm cell line (1047.2)
•  Induction of hypoxia-inducible factor-1α overexpression by cobalt chloride enhances cellular resistance to photodynamic therapy

Great suggestions on this tag and for a list of Inhibitors of HIFs/Hypoxia Signaling as well as for technical answers to than 20 similar/most frequently asked questions on hypoxia/HIFs, I would suggest reading: http://www.novusbio.com/support/hypoxia-and-hif-faqs

using of cocl2 does not have other side effect on your cells? I think if you want to evaluate hole effect of hypoxia on your cells don't use cocl2! because HIF 1 Is not the only downstream target of hypoxia!

The PetakaG3 bioreactor system is the only way to go. It is made by Celartia (Ohio). It so simple to use, you get great results, it is relatively cheap and does not even require a CO2 incubator.  Furthermore, you can send the plate anywhere in the world and the cells will remain viable. I can not believe that we are still using old tissue culture techniques when we know that it does not represent what is happening in the body. Why do scientists continue to use outmoded and inappropriate techniques when we know better. Very perplexing. It is ignorance or laziness. We are suppose to be scientists and not herd followers. 

The induction by CoCl2 is not the same as hypoxia. The Petaka plate is self regulating and does not require CO2. Just Google The Celartia blog and you will see videos on how to use it. If you need individual help just email me at [email protected]. Again, this system is affordable and require little training. Regular tissue culture at 20 percent oxygen is just plain wrong! Just because you get data does not make it meaningful. The body operates in the 5 percent oxygen range. We are wasting precious resources by doing non physiological science. Trying to do drug studies in vitro at 20 percent is wrong. Just thing about it and you will realize that I am right.

by using hypoxia chamber (life technologies)with (mixture gases 2% O2, 5% Co2, 93% N2 ) or 0.5 mM DMOG treatment to the media.

Article Induction and Testing of Hypoxia in Cell Culture

I wrote an article that you may help you consider the measurements your next hypoxia chamber or workstation uses. It compares chambers that use absolute measurements vs those that use relative ones. When you do go out and look for a chamber it should inform you to some of the questions you may want to be asking. Give it a read here:

https://www.linkedin.com/pulse/bringing-hypoxia-chambers-21st-century-making-case-absolute-croft/

I would suggest looking into the newer workstations out there that can correct for the daily changes in the barometric pressures (which can cause havoc on the oxygen levels cells actually "see") and allows for control of O2, CO2, temperature and humidity. Also as others have noted on similar questions any chamber should have some form of HEPA filtration to clean the internal air.

An example of a chamber like this is the HypoxyLab incubator and workstation. Hope this helps.

https://www.scintica.com/products/oxford-optronix/hypoxia-workstation/