Adhesion problems Balb-3T3 - (Mar/07/2006 )
I have a question about 3T3 cells. We are performing a photo toxicity test and everything goes okay until the washing procedure. Our cells are attaching okay to our 96-wells plates, but after washing we are losing cells. There is a wide “ring” where we lost our cells. First there is a ring with cells and then without and in the middle of the plate the cells are also still attached. This is the same for every well. We thought it had to do with the plate (we use Falcon plates), so we tested other plates, but than the loss of cells is even worse.
I have searched the internet for information about this, but I can’t find anything that can help us. I thought we might have to use a special coating, but the same here, I can’t find any information about it. And random testing of 96-wells is not doable because of the wide variety of 96-wells plates and different coating-layers.
Does anybody know if we should use a special coating or have other suggestions? All ideas are welcome.
Not even a hint? Or a link to information about photoxicity assay's?
I'll keep searching....
Although I've never worked with Balb-3T3, I suggest you try to coat the plates with materials such as gelatin or collagen. I use these for my HEK293 cells. It makes them adhere very nicely and I don't dislodge them while washing....
If this does not help, try coating the plates with poly-lysine
We'll try that, maybe it will work.