( Conclusion ...)Cell Proliferation Assay Using the Benchmark Plus Microplate Spectrophotometer

Conclusion
The measurement of cell viability and cell division is used in a wide range of experiments to determine growth factor activity, for drug screening, and to determine the effects of chemical transfectants on human cells. We tested the Roche cell proliferation kit for measurement of cell density of A549 cells grown in 96-well plates. After 24 hr, when the cells were ~90% confluent in the wells with the most cells, we performed the cell proliferation assay.

At low cell density (cells ~50% confluent on the plate), there was a proportional increase in OD 475 with increasing cell number. At high cell density, the increase in OD 475 was not proportional to the increase in cell number. As seen in the figure, this increase was independent of the length of incubation with the reagent between 1 and 3 hr. However, at shorter incubation times (0.5 hr, data not shown), the OD 475 was proportional to cell number up to 5070% confluence.

Bibliography
Guide to Cell Proliferation and Apoptosis Methods, Roche Applied Science, available online at http://www.roche-applied-science.com/fst/publications.htm?/ prod_inf/manuals/cell_man/cell_toc.html

Roehm NW et al., An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT, J Immunol Methods 142, 257265 (1991)

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