McLeod, J. D.
Inter-laboratory study of the in vitro dendritic cell migration assay for identification of contact allergens.
Toxicology in Vitro, 25 (8).
Publisher's URL: http://dx.doi.org/10.1016/j.tiv.2011.09.021
The aim of this study was to investigate the transferability of technology and reproducibility of MUTZ-3
derived Langerhans Cell (MUTZ-LC) migration assay. The protocol was transferred from the NL-lab to two
Sens-it-iv project partners (UK-lab, Italy-lab). Intra- and inter-laboratory variation with regards to MUTZ-
3 progenitor culture, differentiation to MUTZ-LC, maturation and migration assay were investigated. In
the transwell-migration-assay, preferential migration of sensitizer-exposed MUTZ-LC towards CXCL12
was observed (three sensitizers), whereas non-sensitizer-exposed MUTZ-LC only migrated towards
CCL5 (two non-sensitizers). Four pre-pro-haptens were also identified by UK-lab. When taking the arbitrary
criteria of at least two of three independent repetitions per laboratory having to have a CXCL12/
CCL5 ratio >1.1 for classification as a sensitizer, all sensitizers tested in all labs were easily distinguished
from all non-sensitizers. The number of repetitions giving false negative or false positive was very low
(only 7 out of a total of 54 repetitions), indicating that both intra- and inter-laboratory variation was
extremely low. Even though only a few chemicals were tested in this study, we show clearly that the
in vitro DC migration assay is transferable between laboratories. The results were consistent between
the laboratories, and the dose response data were reproduced in the three laboratories.
|Uncontrolled Keywords:||in vitro models, immunology, EU|
Professor J. Mcleod
|Deposited On:||12 Jan 2012 11:24|
|Last Modified:||02 Dec 2013 06:00|
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