Adherent cell culture can be an extremely delicate process, and even with the right techniques and tools it only takes a small problem to upset your culture. The sooner you can recognize any signs of stress or even contamination, the easier it will be to pinpoint the source of the problem and remedy it.
To isolate the problem, it’s a good idea to get into the habit of checking cell health each time you examine your cells for confluence. The good news is that this doesn’t have to be time-consuming, and to make it even easier we’ve created a short checklist with some of the most important things to look out for. In many cases, it’s as simple as 1- 2- 3.!
- Are some cells starting to detach from the substrate? In adherent culture, cells bind to the substrate or ECM, which function as the cell’s regulators. Without these, cells may lack the essential nutrients and structural support they need. The phenomenon of cell detachment usually indicates that the cells need to be sub-cultured or passaged.
- Can you see the formation of several large vacuoles? If you can, this might be a sign of cytoplasmic vacuolation in response to cellular damage. Vacuolation often occurs due to mycoplasma contamination, which can result from poor aseptic cell culture and media filtration techniques. Another reason behind vacuolation could be the cells being cultured too densely, leading to intercellular competition for nutrients.
- Are there granules around the nuclear membrane? These may occur as a result of repressed translation due to oxidative stress, heat shock, UV exposure or even viral infection.
None of us has unlimited time to keep constant watch over our cultures so it becomes vital to be able to spot these irregularities early on. Checking for these tell-tale changes in cell morphology means you’re more likely to be able to rescue or restart a faltering culture in the initial stages, potentially saving your experiment, time, budget, or all three!
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