We've been working with C2C12 cells, and we've consistently encountered a recurring issue. In some of the wells, the cells are dying out. We've tried various approaches, including different pipetting methods, alternative media, and even changing the CO2 incubator. We also ensure that our medium is kept warm. While this problem typically occurs in the last row of the plate, it can also manifest in other parts of the plate. Despite trying a range of solutions, we are currently at a loss for a solution.
I have attached photos of both healthy and dead cells.
Hello Kamil Klejbuk!
I'm going through the same problem as you. I have been working with this cell for more than 8 years and I have never been through this. I thawed all the vials, and this happens right after freezing, or after differentiation, the myotubes remain and the myoblasts are the first to appear dead. I have already tested redoing all the solutions, such as culture medium and buffer, in other greenhouses, several culture bottles and brands. The only thing, that we have had problems previously with adhesion, and death, and even changing, we continue to use Gibco's fetal bovine serum. At the time, the problem was that it seemed that there was something in the fetal bovine serum similar to trypsin, or the lack of some nutrient, which had a similar effect. So much so that this same problem happened in different ways, in other cell types that I culture, such as HepG2 and 3T3-L1 fibroblasts.
This week I will perform a test by thawing another vial of C2C12, but with different brands of fetal bovine serum. I come here to bring information.
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