Our CO2 tanks run out of CO2 so our cells in the incubator spent a whole night (from 4 p.m. yesterday to 10.a.m. today) under very low CO2 concentration. The media of course turned to fuchsia and a part of the cells are dead now. Now we have new tanks applied and the CO2 is increasing.
What should we do with the cells in the incubator? Should we restart all of the experiments? Or can we use the surviving cells?
Thanks.
Usually it should be okay if you have not opened the incubator door several time after 4PM to 10AM. If you think some cells are dead then you subculture them for couple of days and then start your expt.
It depends on several factors, like the cell types used and the requirements of an experiment, i.e. how much are your experiments dependent on the happiness of your cells - for many experiments, it might be OK to use recovered cells; I guess, that this happens everywhere, although there should be a beeper warning for low CO2 pressure in the incubator. Some cells appear to be quite resistant to ph-changes, some are definitely very sensitive; in other cases the pH can influence the characteristics of what one wants to measure, for example, cell adhesion receptors like integrins. I've seen biophysicist handling cells quite wrong, i.e. performing experiments with sensitive cells over many hours out of a CO2 incubator and measuring ph-sensitive adhesion receptors like integrins (and even at room temperature). I wonder how such labs still can publish in Science regularly. I developed a more expensive approach and even adapted many cell cultures to CO2-independent media (and of course using higher, close to physiologic temperatures) to measure (to the best of my knowledge today) for the first time single-molecule adhesion events on living mammalian cells under close to physiologic conditions.
One may expect that some surviving cells from cell clones can represent different subclones, but any cell culture system selects for some sublcones, that's why after several passages in different labs, cell lines can appear differently and differ from the early passages...
What might be worse is to store frozen cells in only one nitrogen tank: I asked biophysicists to store my unique cells in two different tanks, to be on the safe side - but they guaranteed there will be technical assistants around to check for nitrogen... of course, those biophysicists killed the relevant cell line.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line