The Björn Ekwall Memorial Lecture by Dr Richard Clothier. “Experience in the Development and Evaluation of In Vitro
Alternative Assays”. Abstract Over the past 30 years FRAME has, through its Alternatives Laboratory (FAL), developed and participated in the evaluation and validation of alternative in vitro methods. During 20 of those years Dr Richard Clothier was the Director of the FAL, and during this time participated in the development of the criteria and methodology required to validate alternative in vitro methods. It has also become clear that transparency is an important feature of in vitro experimental methods, including data acquisition and analysis. From the outset of participation in inter-laboratory studies it was considered that human cells should be used in vitro to evaluate potential human adverse reactions. FRAME has also been aware of the problems that have been raised in terms of in vitro assays, including a) should animal or human cells be used?, b) should primary cells or cell lines be used?, c) should monolayers of 3D models be employed? and d) how and with what should the in vitro data be compared to achieve the best human prediction of adverse effects? This presentation considers the experiences in the FAL and reflects upon these issues and how FRAME and the FAL have tackled them and learned from them.
Biography Dr Richard Clothier is a FRAME Trustee, and before he retired was Director of the FRAME Alternatives Laboratory (FAL), at the University of Nottingham medical school. He initially worked on tumour induction in amphibians, but they proved very difficult to induce (the South African clawed toad appears highly resistant to cancer). With Prof. L Rubens at Reed College, Portland, Oregon, USA he studied the mechanisms whereby these animals were so resistant to tumours. One aspect of his research was to culture tissues in vitro, initially from amphibian, then later in vitro culture and toxicity testing with mammalian cells, in collaboration with Prof Michael Balls. Dr Clothier took over directorship of the FAL in 1993. During this period the FAL’s research focused on development of in vitro alternative assays for toxicity. It also participated in the evaluation, via national and international blind trials, of the relevance and reliability of the in vitro approach to acute toxicity prediction. The FAL was involved with the FRAME study in 1983-6, the EC/HO and COLIPA on eye irritancy, the MEIC, SDA, ECVAM/ICCVAM, EU/ COLIPA Phototoxicity, and the AcuteTox studies. The FAL has been involved with the development of FRAME Kenacid Blue assay, the Neutral Red Release assay, the Fluorescein Leakage assay, human skin model for Schistosoma infections, innervated corneal epithelial models and bronchial models for prediction of squamous metaplasia. Dr Clothier was an Associate Professor in the School of Biomedical Sciences in the University of Nottingham, where he taught anatomy and histology, and was the departmental safety officer. He retired in 2005.