tldr: I have a dry well of a 24 well plate that did have suspension CHO-K1 cells in it up until about 5 days ago when I upscaled them. The plate has been kept in a humidified incubator since (a few of the other wells still have cells in them- hence why I didn't bin the whole plate). I can still see cells in the dry well under the microscope and have now topped up the well with medium. What are the chances that they will recover from this? Could the small amount of residual liquid and humidity have been enough to keep them alive?
Background: I'm in a bit of a tricky situation because I have been trying for months to make a point mutation in CHO-K1 suspension cells using CRISPR. I'm nearing the end of my PhD so I don't have enough time left to repeat. I had about 50 clones to screen and 1 of them came back positive for the mutation with no signs of it being polyclonal. However, my colleague knocked the 6 well plate containing these cells over in the incubator. The cells were in the front 3 wells of the plate and it fell onto its back edge, causing liquid to spill from the front wells into the back wells and some spilled out completely. I'm about 90% confident that no cross contamination occurred in the front wells (I even recreated the spill using a plate full of water to see what happened). The cells have survived the ordeal but I obviously need to sequence them again to make sure they weren't contaminated and I am cloning them out a second time but this will delay me by a few weeks that I don't really have. Most of my hopes are on the 24 well plate cells recovering. Has anyone had any experience reviving cells like this? Please let me know!
One last note- obviously this situation has been a huge wakeup call to me to keep backup plates/flasks of all my cells in future! Hindsight eh?
Hello
It depends upon the cell's ability to handle stress to which it is exposed. Depending on the level and mode of stress a number of defense mechanisms and pro-survival strategies are undertaken by the cell, and when these fail programmed cell death is activated.
Deprivation of nutrients to these cells could induce self-digestive process as an intracellular energy source before undergoing apoptosis. The cells undergo morphological changes and start shrinking.
How do the cells appear under the microscope? Are they normal? The chances of recovery of these cells are less. Nevertheless, you just need to wait and watch.
Hope for the best.
The below book can help you
https://www.vanderbilt.edu/viibre/CellCultureBasicsEU.pdf
I don't know the answer- and you probably know by now how it went. CHO cells are pretty sturdy work horses so if any cell line could take it would be CHOs. I wasn't sure if the well was completely dry? I hope they made it or things are sorted. Solidarity though- last few months of phd can be very stressful.
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