Comparison of Cell Viability, Proliferation and Apoptosis Assays

Cell-based assays are widely used in basic and translational research as cost-effective and accessible models to mimic in vivo responses. To obtain reliable data, assessing the health of cultured cells prior to any assays is essential. Furthermore, many cell-based assays require quantification of cell growth. Cell health and growth can be determined by quantifying cell viability, proliferation, or apoptosis. Below, we compare some commonly used assays to help you determine which type is suitable for your experimental design.

  • Cell viability assays enumerate the ratio of live and dead cells in a population. Cell viability can simply be achieved by staining and counting live or dead cells.  A deeper assessment of cell health can be attained by measuring cell metabolic activity, such as the ability to reduce tetrazolium salts in MTT and WST-1 assays.
  • Cell proliferation assays assess dividing cells. Some common assays include BrdU incorporation (BrdU Assays) that can directly measure DNA synthesis, the detection of proliferation markers such as MKI67, proliferating cell nuclear antigen (PCNA) and TOP2B, or the indirect measurement of cell metabolic activity (MTT and WST-1 assays).
  • Apoptosis assays are employed to evaluate the extent and timing of apoptosis by measuring key cellular events in programmed cell death, such as DNA fragmentation in TUNEL, GSH depletion, and caspase-3 activation.
  • In addition, there are assays that can facilitate the assessment of cell health, such as cell senescence assays that detect senescence markers like senescence-associated β-galactosidase (SA- β-gal), and LDH cytotoxicity assay that can measure cell membrane integrity.

Table 1. Commonly used assays for evaluating cell growth and health.

Assay Name Cat # Viability Proliferation Apoptosis Other
Live/Dead Cell Staining 8138 x
MTT Assay 8028 x x
WST-1 Assay 8038 x x
TUNEL Assay 8088 x  
Caspase-3 Assay 8228 x  
GSH/GSSG Ratio Assay 8558 x  
Cell Senescence Assay 8058     x
LDH Cytotoxicity Assay 8078   x

Table 1 summarizes the most commonly used cell viability, proliferation and apoptosis kits.

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