If I am using growth medium containing BPE instead of FBS, is it still necessary for me to serum starve my cells for 24 hours before subjecting them to drug treatment? My purpose is to study cell inflammatory reactions and to collect the final cell culture supernatant for ELISA assays. I'm culturing normal human keratinocyte cells.
Hello Phylia Chew
Serum starvation will be required to achieve cell synchronization before you start the drug treatment. Elimination of serum from culture medium removes knowns and unknowns, reduces analytical interference, and provides more reproducible experimental conditions. Moreover, it reduces basal cellular activity and makes the population of proliferating cells more homogenous, since they withdraw from the cell cycle to enter the quiescent G0/G1 phase. You should carry out serum starvation-induced synchronization, followed by re-stimulation of cells with bovine pituitary extract (BPE).
For your experiment, I feel you should starve the cells for 24 hours to achieve synchronization. The cell culture medium may be changed after 24 hours to serum-free medium supplemented with BPE to stimulate cellular proliferation for a brief period (say 3-4 hours). After 3-4 hours, you may subject the cells to drug treatment.
Best.
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