Computerized microassay of keratinocyte cell-plastic attachment and proliferation for assessing net stimulatory, inhibitory and toxic effects of compounds on nonimmortalized cell lines.
Skin Pharmacol
; 6(2): 135-47, 1993.
Article
em En
| MEDLINE
| ID: mdl-8352951
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Queratinócitos
/
Adesão Celular
/
Contagem de Células
/
Divisão Celular
Limite:
Humans
Idioma:
En
Revista:
Skin Pharmacol
Assunto da revista:
DERMATOLOGIA
/
FARMACOLOGIA
Ano de publicação:
1993
Tipo de documento:
Article