Evaluation of a proposed in vitro test strategy using neuronal and non-neuronal cell systems for detecting neurotoxicity.
The European Commission White Paper, "Strategy for a future chemicals policy" (EC, 2001) is estimated to require the testing of approximately 30,000 "existing" chemicals by 2012. Recommended in vitro tests require validation. As the White Paper (EC, 2001) requires neurotoxic data, this study evaluated an in vitro testing strategy for predicting in vivo neurotoxicity. The sensitivities of differentiated PC12 cells and primary cerebellum granule cells (CGC) were compared to undifferentiated PC12 cells which can indicate basal cytotoxicity. Cytotoxicants and neurotoxicants selected for testing covered a range of mechanisms and potencies. Neurotoxicants were not distinguished from cytotoxicants despite significantly different cell system responses using all endpoints; cell viability/activity, ATP depletion, MMP depolarisation, ROS production and cytoskeleton modifications. For all chemicals tested, neuronal-like cell systems were generally less sensitive than undifferentiated PC12 cells. Acute oral rodent LD(50) values correlated with cytotoxicity IC(50) values for the respective chemicals tested in each cell system. This study concluded that although simple non-specific assays are required to distinguish basal cytotoxicity from specific neurotoxicity by using different cell systems with different states of neuronal differentiation, further work is required to determine suitable combinations of cell systems and endpoints capable of distinguishing neurotoxicants from cytotoxicants.
Gartlon
J
J
ECVAM, European Commission Joint Research Centre, Via E. Fermi 1, Ispra 21020, Italy. jgartlon@hotmail.com
Kinsner
A
A
Bal-Price
A
A
Coecke
S
S
Clothier
R H
RH
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001562:Benzimidazoles; D002232:Carbocyanines; D002460:Cell Line; D002469:Cell Separation; D002531:Cerebellum / Q000166:cytology; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007202:Indicators and Reagents; D007928:Lethal Dose 50; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects*; D020258:Neurotoxicity Syndromes / Q000473:pathology*; D009498:Neurotoxins / Q000633:toxicity*; D010078:Oxazines; D016716:PC12 Cells; D051381:Rats; D014966:Xanthenes
D001562:Benzimidazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D007202:Indicators and Reagents; D016900:Neurofilament Proteins; D009498:Neurotoxins; D010078:Oxazines; D014966:Xanthenes; C005843:resazurin; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; D000255:Adenosine Triphosphate