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.
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.