I am working on a project in which we obtain lymphocytes through Ficoll density gradient separation and then culture them for ~3 weeks. Naturally, we add IL2 to our cell culture media. There are two practices that I have seen students do: one is to make a big batch of media with IL2 in it at the start of the experiment and use it over the 3 weeks (The media bottle is stored in the fridge at 4 Celsius. The second is to make the big batch of media without IL2 and only add IL2 to the culture flask when the media needs to be changed. Which approach do you think is better? (Note: in the second approach, the aliquots of IL2 undergo repeated freeze-thaw cycles. Since very little concentration of IL2 is needed in the culture (ng/mL), we do not make single-use aliquots as there would be a lot of them. We make aliquots with 100ug/mL concentration, thaw them upon need, use whatever is needed, and then freeze the rest for later use.)
Hi Mubasher Iqbal
Here are a few advantages of preparing aliquots and using IL-2 freshly and how we do.
* Minimizes repeated exposure of IL-2 to air and light: This can degrade the protein and reduce its activity.
* Ensures consistency: Preparing single-use aliquots ensures consistent concentrations for each experiment.
* Reduces waste: Avoids discarding unused portions of reconstituted IL-2.
It is a bit time consuming but reduces IL-2 degradation and wastage, since it is very costly. In case you are using regularly and storing at proper temp and not exposing to air and light, you can always go ahead with preparing it along with media.
Thanks,
- Badges
- Science topic
- Similar topics
- Chemistry
- Pharmacology
- Pharmaceuticals
- Drugs