J774.A1 marophages grow well in flasks but not in 24-wells plates - (Jun/10/2014 )

Hi all,

I'm doing phagocytosis assay using J774.A1 macrophages. I've had no problems growing them in both cell culture flasks and 24wells-plates until now. For the last several months the cells would grow fine in flasks but every now and then when seeded into 24-wells plates they would detach from the bottom, and some of them even have "necrosis-like" cell lysis (even before infection). I used DMEM+10%FBS+L-glutamine+antibiotic as growth medium. Any suggestions would be very helpful to me.

Thanh

Does it happen every time when you plate them on 24 wp these days?

If so, perhaps it's time to defrost a new batch of cells?

Also, the plastic on the flasks and plates could be slightly different, so the cells are less adherent to plates. Have you, by any chance, switched to a new source of plates recently?

If this problem only happens occasionally, perhaps it relates to the particular way you do the passaging? I don't know if you need to trypsinize these cells or just scrape them, but could it be related to overtrypsinization? Or any other small difference in the procedure?