Here - Stiftung Forschung 3R

were also evaluated as a way to overcome
their intrinsic limitations by generation
of metabolically competent cell lines.
Furthermore, in vitro models were evaluated
regarding their metabolic capacity and
the possible use of these models for
generating data on liver clearance for
PBBK-TD modelling (in collaboration with
WP5). In addition, metabolite formation of
selected compounds in vitro was analysed
and compared to in vivo literature data
as well as to predictions made with the
METEOR software.
The ultimate goal of WP7.3 was to provide
quantifiable information that could be
integrated in a wider assessment of in
vitro cytotoxicity that could anticipate in
vivo toxicity of chemicals. Metabolically
competent cells (rat hepatocytes), noncompetent
hepatic cells (HepG2), and
non-hepatic cells (3T3 mouse fibroblasts)
were used to investigate the effects of a
selected list of reference compounds. In
WP7.3, cell systems suitable for hepatic
transport assays of anions bile acids
and/or xenobiotics, including doubletransfected
cells and ATP dependent
transport systems were also generated.
Furthermore, pilot experiments using
the newly developed fluorescent bile
acid derivatives and cell systems were
performed to determine their robustness
and suitability medium-throughput
testing.
WP7.1: Alerts & Correctors in
Toxicity Screening (III): Neurotoxicity
Acute toxicity may be a result of impaired
neuronal function, either in the peripheral
or the central nervous system. The
overall objective for WP7.1 was to carry
out testing of the ACuteTox reference
chemicals in an optimised neurotoxicity
test battery, according to well defined
SOPs, and to deliver high-quality in vitro
data. The selected assays were identified
as the best performing assays out of a
larger set of assays in which 16 general
and 10 neurotoxic reference compounds
were tested. The criteria for selection
were the ability of the neurotoxicity
assays to (i) identify ‘neurotoxic alerts’,
i.e., indicating alteration of neuronal
function at lower concentrations than the
general cytotoxicity indicated in the 3T3/
NRU assay (see WP2), and (ii) ‘correct’
underestimated toxicity as determined
by the 3T3/NRU assay. Eight different
cell models for the nervous system were
used for the studies on approximately
70 endpoints; in pure enzymes, native or
differentiated human neuroblastoma SH-
SY5Y cells, primary cultures of mouse
or rat cortical or cerebellar granule
neurons, mouse brain slices and mature
re-aggregated rat brain cells. Several of
the endpoints were analysed in more than
one cell model.
WP7.2: Alerts & Correctors
in Toxicity Screening (IV):
Nephrotoxicity
The kidney is especially susceptible to
toxicity because of its role in excreting
compounds, which involves a high blood
supply, concentrating, metabolising and
transporting compounds. The focus in
WP7.2 was on developing in vitro assays
that reflect the role of the kidneys in vivo
PROGRESS REPORTS FROM EU-FUNDED PROJECTS
Progress Report 2011 & AXLR8-2 Workshop Report
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