3 replies to this topic

[wzolla86]

Hello,

This is my first post.  This looks like a very helpful forum.  Anyway, this morning I started warming up my media components for my Vero cultures (medium 199, Bovine Calf Serum, and 200nM l-Glutamine).  I thought I had set the temperature to 37ºC, but I guess I didnt lock the temperature in and it started warming up to the default 60ºC.  I cecked back and hour later and the temp was up to 57ºC!  I took them out and stuck them in the fridge.  Any chance that these are still salvagable?  I know the BCS is probably just heat-inactivated by now, but I'm mostly worried about the l-glutamine and medium 199.

Thanks!

Will

Edited by wzolla86, 17 August 2012 - 05:23 AM.

[bob1]

Throw them out, it's not worth the risk that something has gone wrong.  The l-glutamine will definitely be stuffed, as will a lot of the other labile compounds.

[oanaotilia]

Hello

Are you sure that glutamine is denaturating at higher temperatures?

[ascacioc]

Amino acids + sugars + high temperatures = caramelization reaction; glutamine will react with the glucose in the media when you heat the medium up. I wouldn't use caramelized medium after 57oC even though it is not as caramelized as when you would autoclave it. On the other hand, I have seen people using autoclaved medium containing both amino acids and sugars at the time of autoclaving, and they've been successful in their experiments. It is not ideal, but it works for them. Anyhow: the correct way is to autoclave/filter sterilize amino acid containing parts and the sugar containing part  separately and then mix them up. Rule valid not only for the mammalian cell culture but also for yeast media and other media containing both amino acids and sugars.

Andreea