Add L-glutamine in cell culture medium? - (Oct/14/2005 )

Please forgive my ignorance, but I have never worked with RPMI before and we will be using some for a cytoxity assay soon...the cells will not be maintained in this medium, it will only be a vehicle for the assay experiment.

Can anyone tell me what is the purpose of L-glutamine in RPMI? Manufacturers sell it with and without, but I am having a difficult time understanding the indications. when would you want it? When is it indicated to use RPMI without it?

Thank you for your help!

A

-aimikins-

Second time I've posted this warning tonight -- I don't do cell culture work...

However, it looks as though some types of cells require exogenous L-Glutamine for normal development in vitro -- see here, for example.

See also this article:

QUOTE

It is now known that a large number of tissues and cells in the body utilize glutamine at high rates and that glutamine utilization is essential for their function. These tissues and cells include kidney, intestine, liver, specific neurons in the CNS, cells of the immune system, and pancreatic ß-cells (see Refs. 11 and 12 for further details)...

...L-glutamine is required for a number of specific biochemical reactions, as outlined above. However, of greater physiological importance to many cells, L-glutamine is a precursor of L-glutamate. This review will highlight the critical role of L-glutamine and L-glutamate metabolism for the maintenance and promotion of cell function in a diverse selection of cell types.

Hope this helps...

-HomeBrew-

If I'm not mistaken (please do correct me if I'm wrong!), L-glutamine is a very unstable amino acid, which is why they prepare media without it and you can add it afterwards. This way, you can store your media for a longer time, and have frozen aliquots of L-glutamine. So, you want to have L-glutamine in your medium if you're using to grow your cells, or have them in there for a longer time.

-vairus-

Hello!

We buy the RPMI in flasks and add glutamine, FBS and sodiumpyruvate to the bottle when we are going to use the medium.
Indeed, some cell lines need the glutamine for optimal growth.

Hoping to have helped you in your decision!

An

-Anneke-

some culture media already contain L-glutamine in the form of "Ultraglutamine", a stable dipeptide (alanyl-L-glutamine)
no need to add more L-glutamine

Seb_

-tryptofan-

thank you all for your replies!

I do not think that I need it for my purposes.

I am growing keratinocytes in KGM, as usual...then I am treating them with an agent for about 30' to 90' and then doing a cytotoxicity assay to see if the agent makes the cells croak. Obviously, there will be media-only controls to rule out any effects from that. But, the media for the cytotoxicity assay must be phenol-red free (colorimetric assay) and we have decided to use RPMI for the 90' treatment prior to the assay.

I don't think the L-glutamine will make much difference for the 30-90' duration of the treatment. But, just in case, a media-only treatment will be done as a control to establish a baseline.

Thank you for all your help!

A

-aimikins-