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

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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

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

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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.

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@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.

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Thank you! Great tip. I’ll get some.

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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.