Hello,

To prevent HEK293 cells from detaching during PBS washes, you can consider using collagen or L-lysine to improve cell adhesion. Here’s an explanation of the characteristics of each:

Collagen: This is a natural extracellular matrix protein that provides a more physiological surface for cell adhesion. It enhances cell binding by simulating an environment closer to in vivo conditions, which can be particularly useful if you’re aiming for more natural conditions. However, it can be slightly more expensive and requires a longer incubation time.

L-lysine: This is a positively charged amino acid that interacts with the negative charges on the plate surface, facilitating cell adhesion. It’s a more cost-effective and quicker option, as it doesn’t require special incubation conditions. However, it doesn’t provide as physiological an environment as collagen.

Both options are effective, but the choice depends on your specific needs. If you’re looking to simulate more natural conditions, collagen might be the better option. If you prefer a quick and economical solution, L-lysine is an excellent alternative.

I hope this helps you decide. Good luck with your experiments!

precoating cultureware with poly d lysine should help them adhere better

Hi,

if you use tissue culture treated plasticware, Hek293 cells should adhere well enough without any coating. Yes, they com off easily but there a a few things you can do:

Don't let them get too confluent.

Add PBS very slowly and not directly to the cells but along the sige or top of your flask.

Do not shake too hared or tap the side of the flask.

Just treat them very carefully, and then it should be fine. We culture Hek cells frequently without any problems.

Best,

Franziska