Induction of Apoptosis - without the use of chemical inducers (Jun/10/2010 )
I am wondering, if apoptosis could be induced in cells via serum starvation or other interventions.
in our laboratory the license to purchase hazardous compounds (e.g. actiomycin D...) is currently outstanding!!
therefore i look for an alternative to induce apoptosis, to have a positive control in FACS analysis!!
thx in advance
UV or any DNA damage, you would have to play with doses and time after dose though.
moljul on Jun 10 2010, 08:32 AM said:
in our laboratory the license to purchase hazardous compounds (e.g. actiomycin D...) is currently outstanding!!
therefore i look for an alternative to induce apoptosis, to have a positive control in FACS analysis!!
thx in advance
serum starvation is possible if cells definitely need supplements to trigger survival; however, I would try to get a chemical inducer to a more defined apoptosis induction (vinblastine, staurosporine etc)
As previously stated, UV is a good non-chemical way to induce apoptosis when working with adherent cells. Find a stratalinker. Remove the medium and the lid from your plate. Type ENERGY, 500, START. This will be a treatment of 50mJ/cm2. Add back some medium (serum free is better). Let the cells incubate in the incubator for at least 8 hours. You will see the appearance of apoptotic cells using the microscope. Apoptotic cells will come off the plate, so when you want to collect your cells just scrape all the cells into the medium, collect the medium, and spin down the cells.