I am isolating fibroblasts from heart tissue. I plate them onto T-75 flask. Usually, yield was always consistent. Since one month, cells start to die either at the end of P0 or after trypsinization. I have checked for mycoplasma contamination, which is negative. I don't see any visible bacterial or fungal contamination. Incubator is also working fine. I use 10% FBS media with 1X P/S antibiotic. Please find the example attached. The isolated cells in my lab, also follows the similar pattern of cell death. I am not able to fix the problem.
Thanks
Chandan
This is intriguing. Have you changed media, serum or plastics recently? a new batch of serum in particular may be high in endotoxin orother agents toxic to the cells. Also a new batch of plastics where cells do not attach properly can cause cell death.
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