hi,all. the cells will be suffered high glucose DMEM (25.0 mM glucose) or glucose-free DMEM (no glucose) 24h, low glucose as control(5.55 mM glucose). However, the serum contain glucose. Should I add serum in DMEM?
I noticed some oxygen glucose deprivation (OGD) experiment using glucose-free and serum-free medium. But serum-free is a harmful stimulation, and induced some gene expression. It may interfere results.
Do u have any suggestion? Thanks!
The answer will entirely depend on the question you are trying to answer. Begin with stating clearly what is it you wish to study and include details.
Hi Shang Fei-Fei,
You can add 5-10% of serum to your cells and have them happy and healthy. As a control I would have cells just with serum; DMEM; glucose free-DMEM; serum+DMEM, and serum+glucose free-DMEM. That is if you have cell to have all this conditions, with this you can have an idea if serum is affecting your gene expression or not. I have done this for macrophages and DMEM alone gives me a lot of cell death, so I found out that i need to add serum to it even if my gene expression changes a little.
Good luck
If the only issue with serum is the glucose, you might use dialyzed fetal calf serum. This is supposed to remove small molecules but retains most proteins. I know Thermo/Fisher sells this in the US, so perhaps you could buy it. If you consider dialyzing some serum yourself, I would recommend using a low molecular weight cut off membrane because (I think) some growth factors are fairly small. Good luck.
Hello Shang Fei-Fei
DEM media by high glucose concentration is indicated or frecuently used in culture cell with hig grade of division and growth. I do not know what is your cell type?
You should use serum as source of growth factors and others essential elements for your cells. Usually the serum concentration is used between 5-10%. A serum-free cell culture media can activate the cellular stress according to the incubation time.
Regards
I agree that you should add serum (possibly dialyzed) to medium. This ensures good health conditions so that the cells can tolerate possible stress due to glucose lowering or deprivation
Hi Shang,
it really depends on what you want to know. In my experience switch from HG to LG DMEM already made a difference and even more so with No Glucose - always with serum, which will add
Dear Shang Fei Fei,
It entirely depends on type of study you have planned.
Serum itself contains certain amount of glucose, which will be given in the serum datasheet. If you add serum, then it cannot be called zero glucose control.
Hope this information will be helpful for you.
Good luck.
Maybe there's also a medium on the marker dedicated for your purpose, like chemically defined serum free medium (like for cultivation of stem cells).
You can also try to deplete the serum from your culture growth conditions gradually, adjust your culture step by step to serum depletion, look it up in the scientific press.
Hi Shang
I think the use of media without serum would be activating stress factors, which otherwise will affect your results and the health of cells. Further, i think you are comparing some effect posed by glucose (either high/ low) and i have seen people performing such experiments in complete presence of serum (10%). Thermo fisher FBS contains less than 50 mg/L of glucose and using the same should not affect your result.
Good luck
Thank you all! Finally I added serum. And I tested the concentration of glucose. As Thomas Mühlenberg said,
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