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

Quantitative determination of DNA synthesis in cell cultures is now a routine procedure in many laboratories. Protocols are available for various applications, especially in cell culture systems. The effects of growth factors, inhibition of cell division by exogenous factors, stimulation/inhibition by cytokines, development of serum-free media, influence of hormones and receptor activity on proliferation, and selection advantages for aneuploid cells in vitro are just a few examples where the determination of DNA synthesis provides important information.

In the past, radioactive tagging of newly synthesized DNA with 3H-labeled thymidine was the most frequently applied method, while today non-radioactive labeling with BrdU, a thymidine analog, is finding increasing use. Although the advantages of non-radioactive immune techniques using BrdU or anti-BrdU are not disputed, the scope and limitations of the available systems for detecting BrdU are not always clearly defined. These practical aspects have been examined in more detail in the experiments described below. Besides enabling microscopic detection at the cellular level, the ELISA technique represents a method for routine screening. Two different detection systems are available: a) the conventional colorimetric test and b) a chemiluminescence assay also based on the anti-BrdU technique. One of these test methods will be suitable for use depending on the problem and the available laboratory facilities.

We are particularly interested in the cultivation of human prostatic epithelial cells through several passages. For this purpose, a special culture medium containing exogenous growth factors and added hormones was developed [1]. In the course of this work we hav
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