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fabrablast primery cell line problem - how to detach cell well (Aug/27/2010 )

Hi readers,
i have prepared fibroblast primary cell line from mouse tail. But while i trypsinized the cells, it's not easy to detache them in 175flask. my procedure is
10ml pbs wash,
5ml EDTA-Trypsin, 5min not detached much, then for 10min, still not detached well.
Thanks for your suggestion!


Do you incubate them at 37 deg C when you trypsinise the cells? If not, try that and see how it goes. Some cell lines can take a while to come off.