Hello everyone, I’m looking for advice on inducing hypoxia in cell cultures without an O₂-controlled incubator.
I would need to maintain a constant hypoxic condition for several hours, and I’d greatly appreciate any suggestions on alternative methods you’ve tried.
Thank you!
Hello Carla Santoiemma,
Alternative method would be to treat the cells with cobalt chloride (CoCl2) to induce a chemical "hypoxia-like" state in cultures. CoCl2 mimics hypoxia by inhibiting the degradation of HIF-1α, which then promotes the expression of target genes associated with hypoxia, such as VEGF. This is a common method to study cellular responses to low-oxygen conditions.
You may use the following protocol.
1. Prepare a 25mM stock solution of CoCl2 in sterile, double-distilled water.
2. Add the CoCl2 stock to your cell culture media to achieve a final concentration, which is typically in the range of 100 to 600uM. The optimal concentration of CoCl2 can vary significantly between different cell types, so it is essential to determine the non-toxic range for your specific cells.
3. Incubate the cells in a standard incubator at 37 deg C,5% CO2 for 16–24 hours, or for a duration that is appropriate for your specific cell line.
Please note that the above method is a type of chemical hypoxia, not a true replication of the entire hypoxic environment. For more accurate and comprehensive studies of oxygen deprivation, especially those involving cell metabolism or long-term effects, a specialized hypoxia chamber or incubator would be more appropriate.
Best,
Thank you very much Malcolm Nobre for the suggestion, I will further investigate this methodology and try to apply it.
A colleague suggested using GENbox microaer, which I understand to be an atmosphere generator employed for microbial cultures.
Do you happen to have any experience with it?
