I would like to know the exact protocol to induce chemical hypoxia in A549 cells. There are some variations that I have noticed in the method in various papers which are confusing. Do we starve the cells 24 h prior to cobalt chloride treatment or we just treat in the serum free media. The recommended and frequently used dose is 100 uM from fresh stock. Please correct me if I am missing something and share the protocol.
hello mam,
serum starvation it self is an condition that changes the epigenetics of a cell, that how it should handle serum starvation eg. c-myc goes down with certain other chromatin remodelers.
its depends how you explain your hypothesis. if you want to mimic tumor niche in cell culture conditions you should take both serum starved and unstarved ;hypoxic treatment.
CoCl2 200uM is good for A549. ref:
1.Prolonged Hypoxia Differentially Regulates Hypoxia-inducible
Factor (HIF)-1 and HIF-2 Expression in Lung Epithelial Cells
2.Inhibition of nuclear factor-κB activity enhanced chemosensitivity to
cisplatin in human lung adeno-carcinoma A549 cells under chemical
hypoxia conditions
both of them (authors) are using 10% serum.
hello mam,
serum starvation it self is an condition that changes the epigenetics of a cell, that how it should handle serum starvation eg. c-myc goes down with certain other chromatin remodelers.
its depends how you explain your hypothesis. if you want to mimic tumor niche in cell culture conditions you should take both serum starved and unstarved ;hypoxic treatment.
CoCl2 200uM is good for A549. ref:
1.Prolonged Hypoxia Differentially Regulates Hypoxia-inducible
Factor (HIF)-1 and HIF-2 Expression in Lung Epithelial Cells
2.Inhibition of nuclear factor-κB activity enhanced chemosensitivity to
cisplatin in human lung adeno-carcinoma A549 cells under chemical
hypoxia conditions
both of them (authors) are using 10% serum.
As suggested by Rakesh a concentration of 200 uM under normal growth conditions containing serum should work in A549 cells (Riddle S et al, Am J Physiol: Lung Cell Mol Biol, 2000). Alternatively, you could use deferoxamine (200 uM) under similar conditions.
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