I am having difficulties in reviving the chicken embryonic fibroblast cell line DF-1. I froze down the cells in complete DMEM medium plus 5% DMSO (half of a entire T75 flask) and thawed following the same procedures as other cell lines. However, even though the cells would attach on the next day, they started to die and float on day 2. I am thinking that the antibiotics (pen-strep) in the culture medium may affect DF-1 growth but shouldn't they only inhibit prokaryotic cell's protein synthesis (as for streptomycin)?? Has anyone come across the same problem?
what was the temperature? less than -256 degrees C??? do it at this temp?
Try to use 10% DMSO next time for cell freezing. And after seeding the cells in flask check cells after every 2 hours and once they settled put fresh and new medium. next day wash cells gently with medium and put fresh medium. It will help as cells are very stressed.
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