Journal
JOURNAL OF BIOMOLECULAR SCREENING
Volume 9, Issue 3, Pages 232-243Publisher
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057103262836
Keywords
ArrayScan(TM); high-content screening; NHDF; proliferation; growth factors
Ask authors/readers for more resources
High-throughput cell-based assays are becoming a powerful approach in the drug discovery process. The ArrayScan(TM) high-content screening (HCS) reader is a cytometer based on a fully automated fluorescence microscope that is able to obtain quantitative information on the intensity and localization of fluorescence signals within single cells over a wide cell population. The aim of this work was to set up an automated HCS multiparameter analysis for the quantification of the in vitro proliferation index of normal human dermal fibroblast (NHDF) cultures. The authors stimulated starved NHDF with insulin-like growth factor-1, platelet-derived growth factor, epidermal growth factor, fibroblast growth factor, or serum, and they quantified the proliferation index by measuring the expression of Ki-67 antigen, the incorporation of bromodeoxyuridine (BrdU), and the phosphorylation of the retinoblastoma protein (pRb). This approach also allowed quantification of the mitotic index by phospho-histone H3 staining and the percentage of cells in the S-phase by BrdU incorporation. The proliferation data from the ArrayScan(TM) assays were validated by comparison with a reference enzyme-linked immunosorbent assay (ELISA) and by flow cytometry. The measured proliferation indices were highly reproducible in repeated measures and independent experiments. The authors therefore propose that the ArrayScan(TM) HCS system could be used for high-throughput multiparameter analysis and quantification of the proliferation of cellular cultures.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available