I'm culturing Chinese hamster ovary (CHO) cells in CD. FortiCHO™ Medium + Glutamax, I'm fallowing "Freedom™ CHO-S™ Kit USER GUIDE For transfection of CHO-S™ Cells (cGMP-banked) and development of stable cell lines for protein production". In this user guide the recommended sub-culturing/passaging density range is 10X10^5 to 20x10^5 cells/mL but what i know is that if i start at cell density 3x10^5 cells/mL it will go upto 70x10^5 cells/ml on day 5 after that viability and cell count drops rapidly due to depletion of nutrients, generally cells are passaged between mid log to late log phase, as my cells reach upto 70x10^5 then i guess 30x10^5 to 70x10^5 would be the mid log to late log phase then why they recommended to passage at 10X10^5 to 20x10^5 cells/mL ?
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