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

e examined the colorimetric and the chemiluminescence variants of the BrdU ELISA (Roche Molecular Biochemicals) in a series of parallel experiments.

Materials and Methods

Details of our special cell culture system and a short review on current measurement techniques for cell proliferation has been published [1-4]. We have investigated the proliferation of two distinct human prostatic carcinoma cell lines, P1/289 and P1/373 previously established in our laboratory.

In preliminary experiments, we had already defined the relevant cultivation and labeling intervals as well as the optimal cell concentrations for these cells. We have now investigated the influence of growth factors (epidermal growth factor, EGF) as well as the effects of insulin (Ins) and hydrocortisone (HC) on DNA synthesis in the cell lines P1/289 and P1/373. For this experiment, 104 cells were seeded in standard 96-well microtiter plates in a volume of 200 l medium/well and cultivated in complete medium (DMEM/F12, Life Technologies) containing 10 % fetal calf serum (FCS) supplemented with EGF (10 ng/ml), hydrocortisone (10 ng/ml), cholera toxin (CT, 10 ng/ml), and insulin (4 g/ml). To examine the effects of the additives (DMEM/F12+) on cell proliferation, the cell lines were also cultivated in parallel in their absence (DMEM/F12-).

A second application concerned another cell culture model: To prevent the outgrowth of fibroblasts when establishing cell cultures from heterogeneous tissue samples, cultivation of epithelial cells in the presence of cholera toxin is recommended. When added to normal cell culture medium, this toxin selectively suppresses the growth of fibroblasts. Accordingly, we cultured human fibroblasts obtained from explants (ZF 07) in microtiter plates. The rate o
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