Normally in cell culture we all always have some back-ground cells death (1-5%). I want to reduce this population (0-1% or low). My aim is to inhibit apoptosis/necrosis/autophagy in my cells for at least two cell doubling time. Do pan-caspase inhibitor will work or Do we have any other suggestion.Can we use combination of inhibitors (cell cycle) to achieve this ?
The term "control" in my question refers to expt set up/readout which will give me the best estimation of cell death (apoptosis/necrosis/autophagy).
It would be helpful if you would indicate specifically about your cell system (murine/human, cell type, primary or immortalized/transformed, and culture conditions).
They are immortalized , non adherent cells, Normal cell culture condition (37 deg , 5% co2). Hope this is sufficient information.
Most immortalized non-adherent cells undergo cell death in part due to the lack of cell survival factors. These factors are normally provided by serum in the culture medium and essential nutrients in the culture medium. Therefore, optimizing culture conditions and identifying cell survival factors could help achieve the goal.
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