This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (30) Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Gasparri, F. Articles by Galvani, A. Search for Related Content PubMed PubMed Citation Articles by Gasparri, F. Articles by Galvani, A. Social Bookmarking                         What's this?

Quantification of the Proliferation Index of Human Dermal Fibroblast Cultures with the ArrayScan^TM High-Content Screening Reader

DRO-Oncology, Pharmacology Department, Pharmacia Corporation, Nerviano, Italy, Department of Biotechnology and Bioscience, University of Milano-Bicocca, Milano, Italy

Mariangela Mariani

DRO-Oncology, Pharmacology Department, Pharmacia Corporation, Nerviano, Italy

Francesco Sola

DRO-Oncology, Pharmacology Department, Pharmacia Corporation, Nerviano, Italy

Arturo Galvani

DRO-Oncology, Pharmacology Department, Pharmacia Corporation, Nerviano, Italy

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.

Key Words: ArrayScan^TM • high-content screening • NHDF • proliferation • growth factors

Journal of Biomolecular Screening, Vol. 9, No. 3, 232-243 (2004)
DOI: 10.1177/1087057103262836


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				J. M. Breier, N. M. Radio, W. R. Mundy, and T. J. Shafer Development of a High-Throughput Screening Assay for Chemical Effects on Proliferation and Viability of Immortalized Human Neural Progenitor Cells Toxicol. Sci., September 1, 2008; 105(1): 119 - 133. [Abstract] [Full Text] [PDF]

				K. P. Sarker, H. Kataoka, A. Chan, S. J. Netherton, I. Pot, M. A. Huynh, X. Feng, A. Bonni, K. Riabowol, and S. Bonni ING2 as a Novel Mediator of Transforming Growth Factor-{beta}-dependent Responses in Epithelial Cells J. Biol. Chem., May 9, 2008; 283(19): 13269 - 13279. [Abstract] [Full Text] [PDF]

				O. Durr, F. Duval, A. Nichols, P. Lang, A. Brodte, S. Heyse, and D. Besson Robust Hit Identification by Quality Assurance and Multivariate Data Analysis of a High-Content, Cell-Based Assay J Biomol Screen, December 1, 2007; 12(8): 1042 - 1049. [Abstract] [PDF]

				L. Rickardson, M. Wickstrom, R. Larsson, and H. Lovborg Image-Based Screening for the Identification of Novel Proteasome Inhibitors J Biomol Screen, March 1, 2007; 12(2): 203 - 210. [Abstract] [PDF]

				F. Gasparri, P. Cappella, and A. Galvani Multiparametric Cell Cycle Analysis by Automated Microscopy J Biomol Screen, September 1, 2006; 11(6): 586 - 598. [Abstract] [PDF]

				R. N. Ghosh, R. DeBiasio, C. C. Hudson, E. R. Ramer, C. L. Cowan, and R. H. Oakley Quantitative Cell-Based High-Content Screening for Vasopressin Receptor Agonists Using Transfluor(R)Technology J Biomol Screen, August 1, 2005; 10(5): 476 - 484. [Abstract] [PDF]

				X. Yu, J. S. Sidhu, S. Hong, and E. M. Faustman Essential Role of Extracellular Matrix (ECM) Overlay in Establishing the Functional Integrity of Primary Neonatal Rat Sertoli Cell/Gonocyte Co-cultures: An Improved In Vitro Model for Assessment of Male Reproductive Toxicity Toxicol. Sci., April 1, 2005; 84(2): 378 - 393. [Abstract] [Full Text] [PDF]

				J. Chen, A. Zacharek, C. Zhang, H. Jiang, Y. Li, C. Roberts, M. Lu, A. Kapke, and M. Chopp Endothelial Nitric Oxide Synthase Regulates Brain-Derived Neurotrophic Factor Expression and Neurogenesis after Stroke in Mice J. Neurosci., March 2, 2005; 25(9): 2366 - 2375. [Abstract] [Full Text] [PDF]