I want to show that my developed nanoparticle targets hypoxia but from doing research it seems like most hypoxic targeting related experiments are done in vivo.
I'd like to be doing in-vitro hypoxia studies that mimics tumor hypoxia condition but not sure which experiments are "most important" to perform.
I would love some insight/suggestions, if possible.
P.S I am using the U87MG cell line.
Hi Nina,
Sharing a paper from JoVE ( you may watch complete experiment on journal page) for induction of hypoxia.
I hope this will help you.
Hi, Nina Gen
You can check HIF-1 expression.
You may add Cobalt chloride to your cell to induce hypoxia in-vitro then check whether your nanoparticle is efficient or not.
Hi,
There are two ways to mimic in vitro hypoxia:
- If you have a hypoxic chamber, you could put your cells inside.
- Otherwise, you may use chemical compound that mimic hypoxia conditions, such as DMOG.
You can actually check HIF-1 expression on western blot or a staining with carboxic anhydrase IX that determine hypoxic activity.
You also could determine the effect of your nanoparticle on cell proliferation and migration/invasion. U87 grow naturally adherent, you could easily perform 2D assays. You also could do these assays in 3D with U87 spheroids using methylcellulose or hanging drop approach.
Hi Nina,
Sharing a paper from JoVE ( you may watch complete experiment on journal page) for induction of hypoxia.
I hope this will help you.
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