Caco-2 adherence at plates - (Aug/22/2006 )
Hi! I´ve been culturing Caco-2 cells on 25 or 75 cm flasks and they had growing ok... But, when I trypsinise to repass to the plates, they have no adherence, getting in suspension. Does anybody has a sucessful experience with growing at plates? Did anybody has already use the TPP plates and got the same problem? Any suggestion is welcome1 thanks! 
I have done plate culture with Caco-2 using Corning plates and found no problem with it.
You should have no problems with Caco 2.2 adherence, in fact they adhere to plastic very firmly. We used to have to wash the cells with both PBS "A" and versene before using the trypsin.
Are you using the correct concentration of Trypsin, or are you over trypsinsing them i.e. for too long a period.
The only other thing is the quality of the plastic. The analogy I always use is that Falcon in my opinion is the Rolls Royce of TC plastic, TPP is the morris minor. The plastic from the different companies are very different and will support cell attachment and growth accordingly. One of the first jobs is to optimise the TC plasticware for every cell.
i use sarstedt - no problems with adhering, i use trypsin EDTA for passaging(Gibco). I find that i get dead patches sometimes when they are differentiating.........no idea why. are the dishes coated for tissue culture or do u have to coat them yourself?
Are you using the correct concentration of Trypsin, or are you over trypsinsing them i.e. for too long a period.
The only other thing is the quality of the plastic. The analogy I always use is that Falcon in my opinion is the Rolls Royce of TC plastic, TPP is the morris minor. The plastic from the different companies are very different and will support cell attachment and growth accordingly. One of the first jobs is to optimise the TC plasticware for every cell.
Yep, I detected two errors at my procedure: 1- the time I was using for Trypsin was too long, because the Trypsin was very old, too. The cells was getting injuried. 2- I´m really gonna do tests with new plates - with new GOOD plates...
thank you!!!