I am trying to treat some compound to see gene regulation signaling in HUVEC. For that I starved HUVEC cells with phenol red free and 0.5% serum and without ECGS media for 1 day. However cells died a lot. When I started cells at ~90% confluent, it reched ~50% cells 1 day after changing the media as above.
Do I really need to use the media (without phenol red and ECGS, and with 0.5% serum) ?
Short answer? Yes. ECGS provides essential growth factors for this cell line. You need it.
What is the medium you use before starvation? Is this just a general basal medium with serum and ECGS, or are more compounds or possibly growth factors added? HUVEC are quite finicky, so just having a basal medium with 0.5% serum is probably not going to keep the alive and attached. If you want a system with more control over the growth factors that are still present upon starvation, you could like into things like EGM2 bullet kits. These already have low serum (2%), so bringing this down to 0.5% for starvation should be possible
EBM-2 Basal Medium might be useful for the serum starvation in your experiments. EBM-2 contains no growth factors, cytokines, or supplements.
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