Detecting Apoptosis / Necrosis with MSC
Detecting Mesenchymal Stem Cell (MSC)
Apoptosis /Necrosis using LUMENESC™
LUMENESC™, like the HALO® and LumiSTEM™ Platforms, all measure proliferation of cells using the same ATP-based bioluminescence signal detection system. Since the initiation of apoptosis requires the presence of intracellular energy in the form of ATP, apoptosis of MSC, CFU-F and cells induced to differentiate into MSC-derived cell types can be detected. Apoptosis can be induced by radiation, drugs and other agents that results in cytotoxicity. The LUMENESC™-96 Research and LUMENESC™-96 TOX Platforms not only detect and measure cytotoxicity, but also apoptosis in the same culture wells without performing a separate assay, e.g. TUNEL, Annexin-V.
To detect apoptosis using LUMENESC™, cultures are prepared in the normal manner. When the plates are processed, instead of taking a single luminescence reading after adding the luminescence reagent (reading A), the plate is left in the luminometer to detect the decrease and nadir of the iATP concentration. At this point, the ATP has been converted to ADP. The HALO® Apoptosis Kits contain an additional reagent that converts the ADP back into ATP. The time at which this converting reagent is added is noted (reading B). As the ADP is converted back into ATP, there is an increase in luminescence followed by a plateau in the luminescence readings. The last reading (reading C) then allows the ADP:ATP ratio to be calculated and therefore the apoptosis. The form of the time course will also indicate whether apoptosis or necrosis has occurred.
