Collagen stability - Old collagen doesnt work? (Jul/27/2007 )

Hi

I am culturing intestinal epithelial cells. It used to be working in my previous lab but it doesnt work at the new lab.
It can attach to the collagen coated plate, but at day2-3, they detached.
I think there are tho possibilities. One is serum lot, and I tried other lot but so far I havent figured it out.
The other is Collagen typeI. It happen to be in the lab, so I used the solution. But it seems old.
The collagen type I is BD's one and from Rat tail. (I used to use Bovine.) The product sheet says it is stable for at minimum of 3 months from day of shipment at 2-8C. (of course it is in fridge). But I thought old collagen is still fine, so I am using it.

Could you tell me old collagen is good enough or how long it is OK in your experience?

Thanks

you could check your collagen by running some on a 5% PAGE gel

you should get 3 nice and tight high molecular weight bands showing that your collagen hasnt degraded.

of course seeing as it is a commercial prep you may also get some other crap in there also!!

good luck

Would Collagen IV extracted from basement membrane or EHS cells be a better substrate than Collagen I, which is usually extracted from bone & tendon matrix? Just curious

The collagen solution you're using, is this liquid collagen from BD or is it one that has been made up from a powder or dilluted from the liquid.

The reason i;m asking is that it is very easy from the collagen I to precipitate out of solution the older it gets, expecially on that has been made up from power or dilluted down from the original stock.

I've not found much differerence between Col I and Col IV for cell attachment, but mostly use Col I as its much cheaper.

I am using BD collagen type I solution, and so far I could see no precipitation in the bottle.
For my cells, I used to compare col-I and col-IV, and there is not so big difference (col-I is slightly better). Of course it depends on type of cells.

I am not sure the quality of collagen is important or not in terms of culture efficiency.
I heard that Vitrogen is high quality collagen, I checked it out, and it is no longer available. Instead of Vitrogen, PureCol is same quality.
http://www.purecol.nu/info.html
I am planning to compare BD collagen typeI and PureCol.

If you have any experience that some specific company's collagen is better than others, please let me know.