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Quantitative Measurement of Cell Proliferation,,,Using the BrdU ELISA: A Comparison,,,Between Colorimetric and Chemiluminescent Detection

etric data as increases of merely 16 % and 20 % were recorded. However, the extent of the cell proliferation already leads to values in the non-linear region. Use of the chemiluminescence technique results in different sets of data. In this case, depending on the cell line, growth advantages of 55 % or 63 % are achieved by supplementation of the medium. The alternative, to choose the linear region of the colorimetric test for the experiments, is in general not very practicable since the problems investigated with these test systems usually involve examination of the effects of several parameters in a single experiment. The positive or negative changes in the proliferation index or incorporation of BrdU may vary from parameter to parameter. Thus, it is often necessary to repeat one or more of the experiments, in some circumstances with differing cell numbers or labeling times.

The colorimetric test can be employed without difficulty in cell systems and experiments in which short labeling times are chosen or in which changes in proliferation are measured at the end of a growth curve, near confluence. This is shown by the results with comparatively slowly growing fibroblasts (ZF 07) and the endothelial cells (HUVEC). In these cases, the two BrdU systems provide almost the same information, namely inhibition of cell division of fibroblasts upon use of cholera toxin or the advantage of serum-free endothelial medium in comparison with a standard cell culture medium. The absorption values of the colorimetric BrdU test are in the linear region; thus the differences in BrdU incorporation can be reliably recorded by instruments and are comparable with those from the chemiluminescence technique.

When measurements have to be made on several differently proliferating cells under different conditions in one experiment, the chemiluminescence
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