Identifying proliferating cells
Discover some of the best methods used to study cell proliferation.
Identifying proliferating cells
Below are some of the best methods used to study cell proliferation. We’ve highlighted in green our recommended techniques for each method type.
For investigating cell proliferation in fixed samples, we suggest using Ki67 because it is well-established and highly cited across both the basic and clinical research areas. MCM-2, another proliferation marker, is steadily gathering data around its use as a prognostic marker in certain cancers, making this something to pay attention to as the research continues. For live cells, EdU is the preferred choice.
Method
Marker
Use and benefits
Limitations
DNA Synthesis
BrdU
- Immunoassay to quantify cells in G1, S, and G2/M
- Trace cell cycle kinetics
- Requires DNA denaturation, impairing co-staining and disrupting DNA morphology
- Complex protocol
IdU & CldU
- Immunoassay to study DNA replication fork progression rates, stability or origin firing
- Two dyes (against IdU and CldU) allow more complex experiments than with a single dye
- Requires DNA denaturation, impairing co-staining and disrupting DNA morphology
- Complex protocol
EdU
- Immunoassay to quantify cells in G1, S, and G2/M
- Trace cell cycle kinetics
- Simple protocol, without DNA denaturation
- Can be expensive
Cellular metabolism
MTT
- Biochemical assay to indirectly quantify proliferating (respiring) cells
- Simple method
- Toxic to cells Insoluble in water – needs to be dissolved in a solvent
- Endpoint measure only
- Metabolic assays may not accurately represent changes in cell growth
XTT
- Biochemical assay to indirectly quantify proliferating (respiring) cells
- Simple method
- More sensitive than MTT
- Sensitivity varies
- Metabolic assays may not accurately represent changes in cell growth
WST-1
- Biochemical assay to indirectly quantify proliferating (respiring) cells
- Simple method
- More sensitive than MTT and XTT
- Metabolic assays may not accurately represent changes in cell growth
Proliferation proteins
PCNA
- Immunoassay to detect cells in G1, S, G2 and M
- Prognostic and diagnostic value in some cancers
- Huge body of supporting evidence
- Scoring is subjective
- Can be less sensitive and specific than MCM-2 in some cancers
Ki67
- Immunoassay to detect cells in G1, S, G2 and M
- Prognostic and diagnostic value in some cancers
- Scoring is subjective
- Can be less sensitive and specific than MCM-2 in some cancers
MCM-2
- Immunoassay to detect cells in G1, S, G2 and M
- Prognostic and diagnostic value in some cancers
- Scoring is subjective