I have been working with cell lines that I have never worked with before, HUVEC and HT29 , the problem started with HT29 then I started seeing in HUVEC also, I would be maintaining the cells throughout the week and even sub-culturing them and they were looking okay with some odd formations (HT29 ->budding or bubble like structures around them) (HUVEC -> long tentacles around some cells).
After the weekend I found most of them dead and detached and the rest are extremely stressed with the budding or bubble like structures around them.
This has happened 3 weeks in a row, I changed the incubator since the first time me and a colleague both found our cells dead after a weekend.
I didn't change the media since we are running low on DMEM.
Is it contamination or one of the changeable factors is causing this?
Hello Yan Yan Zhao,
Budding, or the formation of protrusions on the cell surface of HT29 cells is a morphological change that can be observed under stressful conditions. It may be a way for these cells to try and alleviate the stress by increasing surface area for nutrient uptake or waste removal, or it could be an attempt to isolate damaged portions of the cell.
For instance, if HT29 cells are allowed to become too crowded (overconfluent), they will experience increased competition for resources like nutrients and oxygen, and waste products accumulate. This can lead to cellular stress, which can trigger various responses, including the formation of bubble like structures.
Similarly, HUVEC cells show the presence of long, tentacle-like structures called filopodia, which are used for sensing the environment. Stressful culture conditions like lack of certain nutrients in culture medium, overconfluent cultures or any other factor that can increase stress can induce changes in cell morphology, potentially leading to increased filopodia.
You need to culture these cells with utmost care so that they do not undergo any kind of stressful conditions. Avoid such things like overconfluent cultures, media not changed, as these cause stress to the cells. Also, use these cells at a lower passage number because with several passages in culture they can develop these kind of structures.
Best,
Yan Yan Zhao , hello.
The cells may be contaminated with mycoplasma. It is recommended to do a test related to mycoplasma contamination. In addition, it may also be caused by cell hypoxia. It is recommended to adjust the culture conditions of the incubator.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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