Something went wrong with our cell incubator over the weekend so to my knowledge the cells inside were deprived of a sufficient level of Carbon Dioxide for about 24 hours. The photo attached shows the state of the cells.
The cells are NTERA-D2 cells. Passage number 16.
Magnification is 10X.
Cells were passaged 3 days ago into a T-25 flask.
My question is; Can I salvage these cells so they can be used for rtq-PCR and colony formation assays? Or should I just thaw a new vial of NTERA cells?
Hello Navodya Silva
Without CO2 supply, cells are adversely affected within five minutes. Usually, when there is a technical fault in the CO2 incubator and CO2 supply is shut, one can minimize pH change by supplementing the media with 25 mM HEPES buffer. Cells can proliferate without CO2 if the medium is buffered with 25 mM HEPES at pH 7.4, but this environment is viable for no more than 10 hours and is highly dependent upon type of cell line and cell concentration.
Cells deprived of sufficient level of CO2 for about 24 hours may be stressful to the cells, and may result in irreversible damage. I would recommend that you should not waste time salvaging these cells as they would not be healthy as before and therefore unsuitable for any cell-based assay. Since you have NTERA-D2 cells in stock, you should go ahead and thaw a new vial.
Best.
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