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problem with embryo carcinoma cell line , p19 , - (Aug/30/2011 )

hi

i am phd student

i am working on p19 cell lines
in the beginning of my first year the p19 cell line was supper confluent nearly 80-90% within 24hr and they were in a good condition
however, nowadays i have problem growing them. they start growing very slowly and they detached from the flask easily.
i have not change the medium alphaMEM with boven calf serum 7.5% and FBS 2.5%

i do not know why they start behave this way

any recommendation or opinion about what may go wrong with p19 cell line

thank you in advance
ohoud

-ohoud-

How long have you had them growing (when did you last thaw a tube)? Did you freeze them in DMSO? Have you tested them for mycoplasma?

-bob1-