3 replies to this topic

[Scott]

Hi all just thought I'd try and get a consensus view on an argument I've been having my laboratory on low and high glucose concentrations.

In cell culture studies into diabetes cell culture medium with a glucose concentration of approx. 25mM is often used and low glucose 4.5mM (which approximates blood glucose levels) is used as a control. My argument is that in a static pool of 4.5mM glucose will be used very rapidly in actively dividing cells and therefore should be considered low glucose. In addition 25mM glucose which is commonly used in most standard media for cell culture conditions would be a better comparison to normal levels as I would suggest that within hours the glucose concentration would be down to approx. 4.5mM in dividing cells.

Frequently I have seen papers whereby people use 25mM as a mimic of diabetes but don't have a flow system for media replacement this seems strange to me as glucose is getting used up rapidly (it is not like culturing cells the cells are just sitting there).

I haven't measured glucose concentrations yet I was wondering if anyone has, also what is everyone elses opion on this.

Cheers,
Scott

[Rutger]

Hey Scott,

to mimic a diabetic situation really well you should add less or none insulin too, cause thats the thing at diabetic patients.
The higher glucose causes trouble, but as you state that will be solved by the dividing cells very quick.

I got diabetes myself and a personal experience, if i have high glucose and add some insuline it runs down in matter of hours.

I dont know what your setup is, but a bioreactor with a greater media compount or adding extra glucose are both things that
may help to keep glucose level higher then cells prefer.

I hope i helped a bit,

Rutger

[epigenetics]

Scott, on Mar 23 2005, 06:58 AM, said:

Hi all just thought I'd try and get a consensus view on an argument I've been having my laboratory on low and high glucose concentrations.

In cell culture studies into diabetes cell culture medium with a glucose concentration of approx. 25mM is often used and low glucose 4.5mM (which approximates blood glucose levels) is used as a control. My argument is that in a static pool of 4.5mM glucose will be used very rapidly in actively dividing cells and therefore should be considered low glucose. In addition 25mM glucose which is commonly used in most standard media for cell culture conditions would be a better comparison to normal levels as I would suggest that within hours the glucose concentration would be down to approx. 4.5mM in dividing cells.

Frequently I have seen papers whereby people use 25mM as a mimic of diabetes but don't have a flow system for media replacement this seems strange to me as glucose is getting used up rapidly (it is not like culturing cells the cells are just sitting there).

I haven't measured glucose concentrations yet I was wondering if anyone has, also what is everyone elses opion on this.

Cheers,
Scott

we change the media twice a week to make sure that the cells we are growing at HG, are always at HG(high glucose), one of our previous lab person did a small expt on this too, the reasult is after 24 hours it was 384 mg/dl and after 48, 341 mg/dl, so, still pretty high. it was repeated in 10 other different samples, got approx same result.
the cells on which the expt was done is skin FB, may be other cells will consume glucose more faster. not sure.
Thanks

Edited by epigenetics, 12 June 2009 - 11:43 AM.

[agador]

You may perhaps engage in a little side experiment... if you're concerned that the glucose concentration of the media appreciably reduced by rapidly dividing cells then it may be interesting to take an aliquot of your media and check the levels in a commercially available glucometer...