Computerized microassay of keratinocyte cell-plastic attachment and proliferation for assessing net stimulatory, inhibitory and toxic effects of compounds on nonimmortalized cell lines

Details

Serval ID
serval:BIB_6BF4948AEB89
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Computerized microassay of keratinocyte cell-plastic attachment and proliferation for assessing net stimulatory, inhibitory and toxic effects of compounds on nonimmortalized cell lines
Journal
Skin Pharmacology
Author(s)
Grando  S. A., Cabrera  R., Hostager  B. S., Bigliardi  P. L., Blake  J. S., Herron  M. J., Dahl  M. V., Nelson  R. D.
ISSN
1011-0283 (Print)
Publication state
Published
Issued date
1993
Volume
6
Number
2
Pages
135-47
Notes
Journal Article
Research Support, Non-U.S. Gov't
Abstract
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.
Keywords
Cell Adhesion/*drug effects Cell Count/*methods Cell Division/*drug effects Cell Line Epidermal Growth Factor/pharmacology Gentian Violet/diagnostic use Humans Keratinocytes/*drug effects Methotrexate/pharmacology Microcomputers Octoxynol Polyethylene Glycols/pharmacology
Pubmed
Web of science
Create date
25/01/2008 16:30
Last modification date
20/08/2019 14:26
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