Document Detail


Microplate spectrophotometry for high-throughput screening of cytotoxic molecules.
MedLine Citation:
PMID:  20447058     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
OBJECTIVES: High-throughput chemical and biochemical technologies are now exploited by modern pharmacology and toxicology to synthesize a multitude of new molecules with bioactive potential, or to isolate them from living matter. Testing molecules in cell systems on a large scale, however, is a rate-limiting step in drug discovery or in toxicity assessment. In this study, we developed a low-cost high-throughput method for first-level screening of cytotoxic molecules. MATERIALS AND METHODS: We used microplate spectrophotometry to measure growth kinetics of human tumour cells that grow in suspension (Molt3) or adherent to the plastic surface of culture wells (HeLa) in standard RPMI medium. Cells were treated with colchicin, idarubicin or paclitaxel under various treatment schedules. The effects were quantified and compared with those measured by optical microscopy using the trypan blue dye exclusion method to reveal dead cells. RESULTS: Proliferation kinetics of tumour cells can be quantified by measuring variations in optical densities of cell samples at 410 and 560 nm wavelengths. For cells that grow in suspension, one single reading at 730 nm may be sufficient to reconstruct growth curves that parallel those obtained by direct cell counting. Effects of the cytotoxic treatments could also be quantified and results compared very favourably with those obtained using standard techniques. CONCLUSIONS: Microplate spectrophotometry is a robust and sensitive method to monitor growth of animal cell populations both in the absence and in the presence of cytotoxic drugs. This method implements existing technologies and can be fully automated.
Authors:
C Tomelleri; C Dalla Pellegrina; R Chignola
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell proliferation     Volume:  43     ISSN:  1365-2184     ISO Abbreviation:  Cell Prolif.     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-05-07     Completed Date:  2010-06-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9105195     Medline TA:  Cell Prolif     Country:  England    
Other Details:
Languages:  eng     Pagination:  130-8     Citation Subset:  IM    
Affiliation:
Department of Clinical and Experimental Medicine, University of Verona, Verona, Italy.
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MeSH Terms
Descriptor/Qualifier:
Animals
B-Lymphocytes / drug effects*
Cell Adhesion
Cell Culture Techniques
Cell Line, Tumor
Cell Proliferation
Colchicine / toxicity
Culture Media
Cytotoxins / toxicity*
Epithelial Cells / drug effects*
Hela Cells
High-Throughput Screening Assays / economics,  instrumentation,  methods*
Humans
Idarubicin / toxicity
Kinetics
Leukemia, Biphenotypic, Acute / pathology
Paclitaxel / toxicity
Sensitivity and Specificity
Spectrophotometry / instrumentation
T-Lymphocytes / drug effects*
Time Factors
Chemical
Reg. No./Substance:
0/Culture Media; 0/Cytotoxins; 33069-62-4/Paclitaxel; 58957-92-9/Idarubicin; 64-86-8/Colchicine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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