I work on HSC-2 and HSC-3 cell lines in a 12-well plate with cell density 1*10^4 and I found a gradual decrease in cell growth over 3 days in the control group given that the passage number is P4. I observed the cells in a 100 mm dish for 6 days to check the confluence level and it kept going down as well. I found some bizarre shapes I didn't know under a light microscope and a contrast phase one. For contamination exclusion, I cultured the cell media on a blood agar plate for 4 days, and no colonies grew. I have attached some photos to help clarify my question.
Good morning,
There can be many reasons for the failure of a cell culture and the problem is frustrating for researchers.
I recommend you take a look at this link
https://www.sigmaaldrich.com/IT/en/applications/cell-culture-and-cell-culture-analysis/cell-culture-troubleshooting
However, you can also find a lot of literature online.
If you look carefully at the images shown in the link below, you will notice that there are also cells with the same morphology as yours.
https://www.merckmillipore.com/IT/it/product/HSC-3-Human-Tongue-Squamous-Carcinoma-Cell-Line,MM_NF-SCC193?ReferrerURL=https%3A%2F%2Fwww.google.com%2F
They may simply be stressed and perhaps need to be planted at a higher density. Or you have a heterogeneous population of cells in which one cell type is prevailing over another.
Test on a blood agar plate exclude the presence of many bacterial strains but certainly does not exclude the presence of mycoplasmas, for example. Check everything: from the appropriateness of the suitable culture medium for your cell type, to the % CO2 in the incubator, to the exclusion of mycoplasma contamination, to the right seeding density for your cell type.
If you can't solve the problem and you purchased the cells from a company, I advise you to claim as soon as possible to have a different batch of cells sent to you.
Good luck!
Thank you for your answer. It helps a lot. So, what do you think of the cells with processes in the HSC-2 cell line?
The tumor microenvironment is very varied in terms of cell types. They could be phagocytes
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