Adherent cells in suspension - (Dec/28/2006 )

Hi!

I'm working with adherent cells, and I'd like to carry out some assays in wich cells are in suspension (not attached), so I was wondering if somebody knows how could I keep adherent cells in suspension for a long periord of time (24 hours or more) and what could I use to avoid cell attachment on culture plates.

Thank you very much!!

this might not help, but why not change to a non adherent cell line? why work with adherent if you need them in suspension?, i can imagine that adherent cells in suspension will act very different comapared when they are attached, they wont grow and their metabolism will also be differ.

it very much depends on the type of cells; epithelial cells f.i. become anoikic but may be you are interested in this;

we use cell culture vials with permanent shaking at 160 rpm for 16h at 37°C to check anoikis

If you culture trypsonized cells on non-treated plates (petri dish, for example) with low Ca++/Mg++ medium (Invitrogen), you can maintain them as aggregates for 24 hrs or more. The cell growth rate is decreased. Another option is to grow them on beads that support cell growth. GE health sceicnes sells these beads.

Hi!

Thank you for your answers!
Actually, I'd like to study anoikis process in my adherent cell line so, this is why I have to find the way to keep cells in suspension. I've been told about covering culture plates with poly-Hema or BSA...Anyway, I'm trying your advices.
Thanks again!

Hi!

Thank you for your answers!
Actually, I'd like to study anoikis process in my adherent cell line so, this is why I have to find the way to keep cells in suspension. I've been told about covering culture plates with poly-Hema or BSA...Anyway, I'm trying your advices.
Thanks again!