co-culture - (Oct/14/2011 )

Hi everybody,

I want to co-culture 2 different cell lines. One will be the cells that present the ligand (feeder cells) and one will be the cells that have the receptors. How do people to co-culturing and be able to distinguish one cell line from the others? I want to do proliferation assay with them, but I am only interested in the cells with receptors and would like the feeder cells to be constant, i.e. not growing. If I do mitomycin c treatment on the feeder cells (commonly used in embryonic stem cell) they will not grow but they will also not produce proteins (or the ligand that I am interested in). Does anybody has an idea of how to make those feeder cells continue to produce the ligand but not proliferating? Any help/idea will be highly appreciated!

Thanks!!

People used to do things like irradiate cells to act as feeder layers. Check out R.I Freshney's Culture of Animal Cells, a laboratory manual.

You could perhaps use transwells?

Thanks Bob1 for your answer!

I was thinking about transwells... But I read that transwells are not optimal when the cell needs to be in direct contact with the other cells. Since the ligand that I am interested in, it is not secreted, it is attached to the cell membrane, so both cells need to be in direct contact with each other.

And about irradiating cells as feeder cells, I am also considering about it. I have done some preliminary research and read that when cells are irradiated their protein synthesis might be impaired. They will either stop or slow down their protein production. So I am afraid that they will simply not produce ligands that I am interested in.

Thanks foe the reference, I'm gonna read it now.