I am generating fibroblasts from healthy donors and used them at early passages. Some of my colleagues lost their cultures after serum starvation prior the start of an experiment. Can overnight serum starvation kill fibroblasts? Can it be donor specific?
Hello Maria Concetta Zarcone
The time frame for serum starvation will vary with cell type and can include anything from 15 - 30 mins up to 48 hours. There are cell lines which can withstand serum starvation for more than 24 hours while others (for instance, primary cultures) may not be able to undergo serum starvation for long hours. Starving cells of serum beyond its capacity may trigger apoptosis and reduce cell survival.
Serum starvation may be carried out in medium with low serum concentration (relative to normal growth medium), typically 0.1–0.5%, but sometimes as high as 1-2% or as low as 0.05%.
Low serum concentration in the culture medium will help cells to remain healthy but not proliferate. For fibroblasts, you will have to optimize the duration of serum starvation. You may initially use 0.1-0.5% serum in the growth media and starve the cells overnight. Check for cell health by recording cell viability and observing the cell morphology. If you are not satisfied with the results, you may alter the serum concentration (low serum) and the duration of serum starvation and choose the best that suits the cells without having any drastic effect on the overall health of the cells (fibroblasts).
Good Luck!
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