1 reply to this topic

[nmyimliuxianzhong]

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!
Leo

[bob1]

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.