I am currently working with H9C2 cell lines. I prepared the complete growth media by using following:
1. DMEM
2. 10% FBS and
3. 1% PS
Finally, I know media (DMEM) by itself is generally stable for 12 months from manufacture when stored refrigerated, and sera is generally stable for 5 years when stored at -20°C or colder.
So, my question is what is exact shelf life of my complete growth media? And, why shelf life of complete growth media is different from DMEM?
The composition of the complete media dictates the shelf life. Some components such as L-glutamine which are essential for cells, degrade within a month. Generally I dont use complete media which is more than a month old.
The rule of thumb is one month in 2-8 degree Celsius because as Kumar mentions, glutamine will degrade. On top of that, each individual item has a different shelf life, you should therefore follow the one with the least shelf life. Sera needs to be stored at -20 degree Celsius, when added to the media it is stored at 2-8 degree Celsius which also shortens its shelf life.
Since your cells are dependent on those nutrients to survive you would want them to be at optimal quality. I am guessing if you store your media at -20 degree Celsius (and do not thaw) it would also increase the shelf life. However, if you are using the media, you cannot freeze and thaw repeatedly, it would defeat the purpose. That is the reason some laboratories working with difficult cell lines only prepare media in small quantities right before having to culture like if you are working with primary cell lines.
We have kept DMEM for month in the cold room and it always worked fine. There is an expiry date on the bottles anyway. But even expired media worked just fine.
P/S and FBS should be kept at -20 for storage and once open in a fridge. You can keep them at 4C for several month.
As far as I remember the PS (I assume you mean PenStrep) has the shortest life time of all components if stored at 4 °C hence this will determine the overall lifetime. You still can use the complete growth media afterwards - lifetime will mainly depend on how often you take it out of the fridge - do you warm up the whole bottle (yes, I have seen this) and how well can you keep sterility. The DMEM lifetime is given for a closed bottle that is constantly kept at 4 ° C.
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