I have procured B16F10 cells from NCCS Pune and maintained them in DMEM with 10% FBS. The cells received were confluent and healthy but after the a first passage, the adherent cells showed stressed morphology with circular surface and black particles. When I tried to trypsinize these cells, they were not getting trypsinized as well. These cells do not grow and nor do they are die and get into suspension. There is no change in the media color or cell confluency. I have been culturing these cells for a long time but such an issue occurred for the first time. I have changed media, trypsin and all other cell culture reagents, but still the issue persists
Hello Dhruvi Shah
B16F10 cell line is very sensitive to environmental changes. Phenotypic heterogeneity and plasticity are traits by which B16F10 may adapt to environmental conditions. They can easily acquire characteristics of melanogenic activation under stress. You mentioned that the cells were confluent when they arrived from NCCS. So, there could be a possibility that these cells must have undergone phenotypic changes due to stress.
Best.
Hey Malcolm Nobre
Thank you for the suggestion but in case of differentiation to melanocytes, they release melanin pigment in the media and media appears blackish. But there is no colour change in the media nor any contamination.
The cells are just in non dividing nor dying phase
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