Hi! The literature (even here in RG) does not provide a clear answer about the best composition for cryopreservative medium to freeze mammal cell lines, specially regarding to the serum component. In your experience, is FBS recommended or not? if yes, which percentage? Does it change between different cell lines or are there standar compositions? Thank you
The best method is to use your growth media with 10-20% FBS plus 10% DMSO.
You can use glycerol instead of DMSO but in my opinion does not work as well.
Cells must be frozen slowly (use an insulated container) at -80oC before transfer to Liquid N2.
Hi Md,
your freezing media is good and I have used it as well, however, it is very expensive if you are freezing a lot of cells. 10-20% FBS in normal medial really does work as well.
Thank you both for your answers. This is exactly the discrepancy I meant: 90-95 vs 10-20% FBS. Do you think that the reason of putting 10 instead of 90% is just to save money? Because, for example, I tried to freeze 293T cells with 95% FBS + 5% DMSO and they all died; but they were well cryopreserved when I did with 20% FBS and 10% DMSO in DMEM
I would propose a simple 20:1 solution of FBS and DMSO (sterile) for cell cryopreservation.
Lucas, your cells died for different reasons.
All the freezing media suggested should work for 293 cells.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line