Computerized Microassay of Keratinocyte Cell-Plastic Attachment and Proliferation for Assessing Net Stimulatory, Inhibitory and Toxic Effects of Compounds on Nonimmortalized Cell Lines
作者:
Sergei A. Grando,
Raul Cabrera,
Bruce S. Hostager,
Paul L. Bigliardi,
John S. Blake,
Michael J. Herron,
Mark V. Dahl,
Robert D. Nelson,
期刊:
Skin Pharmacology and Physiology
(Karger Available online 1993)
卷期:
Volume 6,
issue 2
页码: 135-147
ISSN:1660-5527
年代: 1993
DOI:10.1159/000211098
出版商: S. Karger AG
关键词: Attachment;Proliferation;Growth stimulation;Growth inhibition;Toxicity;In vitro assay;Keratinocytes
数据来源: Karger
摘要:
Testing of pharmacological agents that affect growth of epidermal keratinocytes (EK) requires a standardized assay. We have developed an assay measuring net effects of stimulatory (e.g. growth factors), inhibitory (e.g. methotrexate) or toxic (e.g. Triton X-100) compounds. The amount of crystal violet staining viable EK attached to the wells of standard 96-well microplates is measured in situ using an ELISA plate reader. Optical density readings are directly converted into cell counts by computer software. Counts obtained by this method strongly correlate with the results obtained using the [3H]thymidine uptake assay and direct cell counts. The assay standardizes measurements of nonimmortalized EK lines with different innate proliferative properties and allows accurate quantitation of EK numbers in the range of 2,500–500,000 EK/well.
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