Cell Proliferation Assays


Cells are structural and functional unit of our body that control and maintain the function of all unicellular and multicellular organisms. The process of cell proliferation and differentiation plays a vital role from the time of embryogenesis to development of whole organism from single‐ or double‐cell embryo and continues its crucial role in maintenance of adult tissue homoeostasis by recycling the old cells with new cells. In addition, abnormal cell proliferation is also associated with various human diseases like cancer. Therefore, cell proliferation assays become crucial to scrutinise the rate of cell proliferation in both in vitro and in vivo conditions. These assays follow different parameters to determine the rate of cell proliferation like rate of deoxyribonucleic acid synthesis, metabolic activity of cells, different antigens associated with cell proliferation and adenosine triphosphate concentration. In the present article, authors have tried to summarise different cell proliferation assays commonly used in laboratories to determine cell proliferation.

Key Concepts:

  • Cell proliferation plays a vital role in regular tissue and cellular homoeostasis for proper growth, development and maintenance of organism.

  • Cell proliferation assays are mainly designed based on three concepts, that is, (1) measuring rate of DNA replication, (2) analysis of metabolic activity and (3) cell surface antigen recognitions.

  • Rate of DNA replication can be analysed by using radioactive or labelled nucleotide analogues, that is, 3H‐thymidine‐ and BrdU‐based assays.

  • Metabolic activity‐based assays include MTT, XTT, WST, resazurin and ATP measurements.

  • Cell proliferation antigen‐based assay targets antigens present in proliferating cells such as markers like Ki‐67, topoisomerase IIB, phosphohistone H3 and PCNA.

Keywords: cell proliferation; cell assays; metabolic activity; ATP; BrdU; MTT assay; rate of DNA synthesis

Figure 1.

Scheme of labelled nucleotide analogue incorporation during DNA replication to measure cell proliferation rate.

Figure 2.

Cell proliferation assay using 3H‐thymidine nucleotide incorporation and radioactive measurements.

Figure 3.

Schematic representation of BrdU incorporation assay into the cell culture system.

Figure 4.

Brief scheme of metabolic activity measurement using MTT and other tetrazolium salts to access cell proliferation.

Figure 5.

Scheme of antigen‐based cell proliferation assay.



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Further Reading

Berridge M, Tan A, McCoy K and Wang R (1996) The biochemical and cellular basis of cell proliferation assays that use tetrazolium salts. Biochemical 4: 14–19.

Fan F and Wood KV (2007) Bioluminescent assays for high‐throughput screening. ASSAY and Drug Development Technologies 5(1): 127–136. doi: 10.1089/adt.2006.053.

Mitchison JM (2003) Growth during the cell cycle. International Review of Cytology 226: 165–258. doi: 10.1016/S0074‐7696(03)01004‐0.

Niles AL, Moravec RA, Eric Hesselberth P et al. (2007) A homogeneous assay to measure live and dead cells in the same sample by detecting different protease markers. Analytical Biochemistry 366(2): 197–206. doi: 10.1016/j.ab.2007.04.007.

Riss TL and Moravec RA (2004) Use of multiple assay endpoints to investigate the effects of incubation time, dose of toxin, and plating density in cell‐based cytotoxicity assays. ASSAY and Drug Development Technologies 2(1): 51–62.

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Yadav, Kamalendra, Singhal, Nitin, Rishi, Vikas, and Yadav, Hariom(Aug 2014) Cell Proliferation Assays. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002566]