title: The FRAME alternatives laboratory database. 1. In vitro basal cytotoxicity determined by the Kenacid blue total protein assay.
abstract: A database of over 280 chemicals has been compiled by using a mouse 3T3-L1 fibroblast-like cell line in exponential growth, exposed to chemicals for 72 hours in a 96-well tissue culture plate format, and determining cell number via the Kenacid blue (KB) assay for total protein. Ranking the chemicals according to their basal cytotoxicity, expressed as the concentration (mM) that inhibits increase in total cellular protein over 72 hours by 50% (the ID50 value) shows a wide range of ID50 values, from 0.00003 mM to 10,096 mM. This information includes the results for MEIC chemicals 1-50, and we have now added basal cytotoxicity data for 23 of the next 25 MEIC chemicals. When the neutral red uptake (NRU) assay was performed with the same cell cultures, before the KB assay, very similar indications of basal cytotoxicity were obtained. Comparisons between the results with 3T3-L1 cells and with a human fibroblast-like cell line, BCL-D1 showed a significant difference in order of magnitude of the ID50 value for only 5 of 52 chemicals. However, there was a difference in ID50 value of more than one order of magnitude for 8 of 24 chemicals tested with an undifferentiated teratocarcinoma cell line, F9.
authors.affiliation: School of Biomedical Sciences, University of Nottingham, Nottingham, NG7 2UH, UK. richard.clothier@nottingham.ac.uk
authors.forename: Richard
authors.initials: R
authors.lastname: Clothier
authors.forename: Elke
authors.initials: E
authors.lastname: Gottschalg
authors.forename: Silvia
authors.initials: S
authors.lastname: Casati
authors.forename: Michael
authors.initials: M
authors.lastname: Balls
mesh_terms: D016475:3T3 Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D000826:Animal Testing Alternatives; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D016208:Databases, Factual; D004305:Dose-Response Relationship, Drug; D006801:Humans; D051379:Mice; D009499:Neutral Red; D009930:Organic Chemicals; D011506:Proteins / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods*; D015262:Xenobiotics / Q000145:classification / Q000633:toxicity
chemical_list: D009930:Organic Chemicals; D011506:Proteins; D015262:Xenobiotics; C069245:kenacid blue; D009499:Neutral Red