.i used 40% fbs for thowing my freezed cell because they 90% dead and i need that 10% remained.and they were so important for me( i had just one stock remained)now i want to know can i trust to genetice background of my cells after this high dose of FBS!!?:(
Although initially high 20-30% serum has been reported to be in use during recovery of frozen cells, FBS cannot completely replace medium because serum is primarily soluble proteins (macromolecules) whereas medium contains well defined composition of amino acids (which is ready to be uptaken and used), salts and glucose.
We found that glucose levels (low, normal, high) can be very critical for certain cell types, especially, e.g. for fast growing cells like HEK293. Cells will spontaneously detach and die when glucose falls below optimal levels.
Therefore, regular maintenance of cells in very high serum level (>10%) is not exactly the preferred or necessarily best method, unless it is something already established in your literature, or empirically determined.
You may divide your cells (e.g. some fractions in small dishes) and culture them in different levels of FBS and see how they perform.
Also, after heat inactivation (HI), the activity of FBS generally declines. Check if your cells are okay with HI.
- Badges
- Science method