An alternative way to determine the health of a culture is to perform a cell proliferation assay, i.e. to determine the number of dividing cells. One way of measuring this parameter is by performing clonogenic assays. In these assays, a defined number of cells are plated onto an appropriate matrix and the numbers of colonies that form are counted after a period of growth. Drawbacks to this type of assay are that it is tedious and it is not practical for large numbers of samples. Another way to analyze cell proliferation is to measure DNA Synthesis. In these assays, labeled DNA precursors (4H-thymidine or bromodeoxy-uridine, BrdU (e.g., Roche Applied Science Cell Proliferation ELISA, BrdU (chemiluminescent) Kit) are added to cells and their incorporation into DNA is quantified after incubation. The amount of labeled precursor incorporated into DNA is quantified either by measuring the total amount of labeled DNA in a population, or by detecting the labeled nucleimicroscopically. Cell proliferation can also be measured using more indirect parameters. In these techniques, molecules that regulate the Cell Cycle (also called proliferation markers) are measured either by their activity (e.g., CDK kinase assays) or by quantifying their amounts (e.g., Western blots, ELISA, or immunohistochemistry).
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