Strong adherent cell lines that can withstand immunocytochemistry and that are good for heterologous expression of proteins?

I’ve been using HEK293 cells to express some plasmid constructs, and I’m frustrated by how easily HEK cells lift up from the surface, even when PDL-coated. Anyone have recommendations for more adherent cell lines that can withstand ICC and that are good for transfection of cytoplasmic and channel proteins? @tberta @liz @thicunha

@achamess Have you tried to generate stable cell lines using lentivirus?? We are currently using. For us, it works well.

It’s been a million years since I did any cell culture, but maybe CHO cells? I used to use those for stable/transient transfection of GPCRs, so maybe they’d also be amenable to channel proteins.

@achamess Our lab always uses CHO-K1 cells for stable/transient transfection of ion channels, they work very well. And they can withstand ICC, too.

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did you try the methanol fixation? I use it for everything including channels. but i’m surprised to hear that HEK cells are detaching from your surface. they are pretty sticky. I use CAD cells that are easy to transfect but a lot less sticky to plastic or glass. It’s still manageable if you want to try it.

Thank you! Great tip. I’ll get some.

I didn’t try the methanol. I did PDL and it wasn’t that helpful but cells were confluent. I’m going to try Cell-TAK. But in general, I find HEK will come off glass or plastic with just medium if too forceful.