I am now do some inflammation study by using RAW 264.7 cells. But there are some problem on RAW cells.
I cultured RAW cells in the DMEM(4.5g/L glucose), 10%FBS,1%P/S. And slitted the cells by trypsin/EDTA. The nitrate oxide in the cultured RAW cells` medium increased during the process of culture. I doubted the effect of trypsin. So I changed my passage way with pippetting. It seemed that the cells had a relatively stable and low nitrate oxide. But RAW cells showed strong reponse on LPS stimulation. And the drug which exihibited inhibition of LPS stimulation in the previous expreiment fail to inhibit the LPS stmulation. And the BAY 11-7082(inhibitor of κB kinase) sometimes showed little effect on LPS-stimulated cells.
I don`t know why. And is there anything wrong with my cells?
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why the drug has little effect on LPS-stimulated RAW 264.7 cellsRAW cells
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