| A novel ex vivo mouse aorta perfusion model (February 2012) |  | 3R-Info-Bulletin 48
includes a report on the completed project
Establishment of an organ ex-vivo tissue slice model for cardiovascular research, in particular for therapeutic atherosclerosis targeting
Prof. Patrick Hunziker, Clinic for Intensive Medicine, University of Basle, Switzerland In this project, aorta tissue of transgenic (ApoE-/-) mice was extracted and cultured ex vivo. A method was developed for obtaining the tissue and its subsequent on-line observation under a fluorescence microscope. It was possible to identify and characterise the sclerotic sections of the walls of the aorta (plaques) following perfusion of the aorta with specific markers. The time taken for changes to the cells could also be determined. The results obtained corresponded to a great extent to findings from studies using ApoE-/- mice, which shows that many studies, for example for preselecting potential new medicines, could be carried out ex vivo.
3R-Info Bulletin 48 | Project 111-08 |
| | New member of the Evaluation Committee (January 2012) |  | On 17 January 2012 the Administrative Board elected Prof. Simon P. Hoerstrup, Head of the Swiss Centre for Regenerative Medicine (SCRM) at the University Hospital in Zurich and at the University of Zurich, to the Evaluation Committee.
Evaluation committee |
| | A new face on the Administrative Board (December 2011) |  | On 15 December 2011 the Administrative Board elected Nathalie Stieger, B. Econ. (St. Gallen), who works for F. Hoffmann-La Roche AG, Basle, to the Administrative Board. She replaces Silvia Matile-Steiner, who resigned from Hoffmann-La Roche.
Administrative Board |
| | Completion of a project (December 2011) |  | An in vitro screening assay for stem/progenitor cells in organotypic hippocampal slice cultures: validation in an experimental model of infant rat pneumococcal meningitis
Prof. Stephen Leib, Institute for Infectious Diseases, University of Berne, Switzerland Cell changes were studied in organotypic hippocampal slice cultures in cell types that might play a role in brain damage and regeneration following bacterial meningitis. The research team succeeded in differentiating the stage of development of neuronal stem/progenitor cells cultured over a period of weeks. The suitability of such cells for transplantation and their behaviour in neuronal tissue were also studied. It was possible to show that the cells interact from a functional point of view with those in the hippocampus tissue slice. This in vitro method could be used for the relevant preliminary investigations and laboratory animals would only be required for the final confirmation of the in vitro findings.
Project 103-06 |
| | 2012 new: Firstly submit a project outline and then an application for funding (December 2011) | | The Administrative Board has decided that in 2012 applications will be dealt with in two stages in order to rationalise the procedure as well as to save applicants spending time on projects that are not likely to be approved for funding.
First of all a project outline should be submitted by 1 February 2012 at the latest.
If the Evaluation Committee considers the project to be relevant to the 3R principles it will then ask the applicant to submit a detailed application for funding.
Call for Grant Applications | Instructions concerning project outlines |
| | 3R methods (November 2011) |  | A new training device for taking blood and administering substances intravenously in rabbits in the form of an artificial rabbit’s ear made of silicon To minimise the animal’s discomfort, students need to practise taking blood from veins and arteries as well as administering substances in rabbits. They can use this newly developed silicon model of a rabbit’s ear for their initial attempts. The model ear is especially useful for compulsory basic courses for people who will be involved in laboratory experiments on live animals.
method M4 |
| | Metabolism as part of alternative testing strategies in fish (October 2011) |  | 3R-Info-Bulletin 47 It is important to be able to determine the possible concentration in fish of lipophilic contaminants that are released into the environment. In order to obtain such information without examining the fish themselves, it is necessary to determine to what extent such substances are metabolised, since this would influence their concentration in the living organism. It has been possible to demonstrate this metabolic processing in freshly isolated fish liver cells using reference substances. By standardising in vitro reports it has been possible to improve the validity of the results obtained using this animal-free method to such a degree that they are comparable with values obtained in fish.
3R-Info Bulletin 47 | Project 108-07 |
| | Revision of the Foundation’s statutes (October 2011) | | On 28 September 2011 the supervisory authorities approved the Foundation’s revised regulations and the modification of the deed of foundation as decided by the Administrative Board on 30 March 2011.
Deed of foundation | Regulations |
| | Publication of Annual Report for 2010 (July 2011) |  | On 30 March 2011 the Administrative Board approved the Annual Report for 2010 on the Foundation’s activities during that year, as well as the financial statements for the same year. A total of Fr. 728,600.00 was paid out for research projects. Seven new projects were approved for funding and the final reports for three completed projects were approved.
Annual Report for 2010 | PDF version |
| | Toxoplasma gondii virulence is predictable in cultured human cells (May 2011) |  | 3R-Info-Bulletin 46
includes a report on the completed project
Evaluation of an in vitro model to identify host parameters associated with virulence of Toxoplasma gondii strains
Dr. Sushila D’Souza1, Pasteur Institute, Brussels, Belgium; 1present address: GSK Biologicals, Wavre, Belgium Toxoplasmosis, a disease which can be acute in man, is caused by Toxoplasma gondii (natural host: cats). The virulence of samples contaminated with Toxoplasma gondii is normally tested on mice. To replace these tests, which cause extreme suffering in the laboratory animals, Dr. D’Souza’s research team have cultured parasites from strains with various known levels of virulence together with human intestinal cells. A correlation between the level of virulence and the number and extent of the foci formation on the cell monolayer was observed, while a reverse correlation was seen with the degree of inhibition of the β-defensin2 proteins, which are responsible for defence against toxoplasmosis, in the intestinal cells. These changes could be used as an indicator for determining the virulence of Toxoplasma gondii without using laboratory animals.
3R-Info Bulletin 46 | Project 107-07 |
| | A new face on the Administrative Board (April 2011) |  | On 30 March 2011 the Administrative Board elected the Foundation’s officers for the coming 4 years. A new person on the Administrative Board is Dr. Markus Schmutz from Novartis Pharma AG in Basle. He replaces Prof. Paul Herrling, who decided to resign owing to his age.
Administrative Board |
| | New project (April 2011) |  | Model development and validation to investigate myeloid cell homeostasis
Dr. Charaf Benarafa, Theodor Kocher Institute, University of Berne, Berne, Switzerland Neutrophil granulocytes play an important role in inflammatory diseases and in the body’s defence against pathogens. They are found only in minute quantities in the blood. In order to investigate their function a large number of laboratory mice, including transgenic types, have normally been used until now. Through the genetic manipulation (Hoxb8) of neutrophil precursor cells the research team intend to obtain functioning neutrophil granulocytes from the precursor cells. These cells will need to be characterised in order to determine that they are indeed behaving in the same way as differentiated cells. If the project is successful it will no longer be necessary to use a large number of laboratory mice nor to breed various types for laboratory experiments.
Project 126-11 |
| | New project (April 2011) |  | Nerve-cell mimicking liposomes as an in vitro alternative for demonstrating the potency of toxins with multistep pathways such as Botulinum neurotoxins (BoNT)
Oliver G. Weingart, biologist, Institute of Food Sciences, Nutrition and Health, Zurich Federal Institute of Technology, Switzerland The registration authorities demand that the level potency be determined for each lot of toxins manufactured from living organisms before they are accepted for medical use. Until now, this has been done using the extremely painful LD50 test on mice. An alternative method involves using in part cell systems, but they are in many cases not sufficiently sensitive and difficult to reproduce. Using liposomes into which the individual steps in the potency pathway have been introduced should make up for this deficiency in the cell systems. It would then be possible to determine the potency level without using the LD50 test in mice.
Project 125-11 |
| | Completion of a project (March 2011) |  | Magnetic Resonance Imaging (MRI) for the non-invasive assessment of lung inflammation and pulmonary function in the rat
Dr. Nicolau Beckmann, Novartis Pharma AG, Novartis Institute of Biomedical Research, Basle, Switzerland To develop treatments for asthma, induced inflammatory and fibrotic changes (early stages of asthma) have been characterised in rats using magnetic resonance imaging (MRI). Compared with conventional pulmonary function measurement and terminal histopathological analyses, the use of this non-invasive method means that the number of laboratory animals required can be reduced by up to 90%, and the condition of the individual animals can be monitored closely. From the point of view of animal welfare, this is a decisive advantage since, if the medication being tested has no effect, individual animals can be withdrawn from the experiment in the early stages of asthma.
Project 82-02 | extended abstract |
| | Completion of a project (March 2011) |  | Non-mammalian Experimental Models for the study of bacterial infections (NEMO network)
Prof. Pierre Cosson,UMC, Faculty of Medicine, Department of Cellular Physiology and Metabolism, Geneva, Switzerland The mechanisms involved in bacterial infections and the cellular defence strategies can also be studied using amoeba and the fruit fly (Drosophila). With this specialised knowledge it would be possible to carry out certain infection experiments using amoeba and fruit flies instead of laboratory animals. In order to promote the use of this method, funding was provided for a 3-year platform for 5 research groups on the 3R Foundation’s website. It is to be hoped that other researchers will use the available models with amoeba and fruit flies for screening potentially effective antibiotics and thus reduce the number of conventional infection studies carried out using rodents.
Project 99-05 |
| | Completion of a project (March 2011) |  | Organotypic CNS slice cultures as an in-vitro model for immune mediated tissue damage and repair in multiple sclerosis
Prof. Norbert Goebels1, Neuroimmunology Unit, Neurological Clinic, University Hospital, Zurich, Switzerland; 1present address: Heinrich-Heine University Düsseldorf, Department of Neurobiology, Germany The processes that lead to lesions in multiple sclerosis are often studied using an animal model (the EAE mouse model). As a part replacement, Prof. Goebels’ research team examined the immunological mechanisms of brain damage in cultured brain tissue slices of transgenic mice. They were able to demonstrate that antibodies and a complementary system cause demyelisation of the axons without damaging them. Antigen-specific cytotoxic (CD8) T-cells did cause collateral damage to the axons, however, but only when the tissue and the cytotoxic T-cells had been activated beforehand. To date it has not been possible to demonstrate this type of damage using the EAE animal model. This could result in a fall in popularity of the animal model and consequently a reduction in the number of EAE experiments carried out.
Project 101-06 |
| | Completion of a project (March 2011) |  | Preparation and evaluation of lipoprotein fractions for the replacement of fetal bovine serum in cell culture media
Prof. Paul Honegger, Department of Physiology, University of Lausanne, Switzerland In most cases cell cultures require media that include fetal bovine serum in order to achieve optimal growth and to maintain cell function. Since the composition of the serum is not defined and varies from charge to charge, and since the acquisition of the serum from unborn calves should be avoided from an animal protection point of view, researchers have long been trying to find a defined serum replacement. Prof. Honegger’s research team have succeeded in demonstrating that, contrary to expectations, a macromolecular protein is not responsible for the stabilisation of cell function in the lipoprotein fraction. The results of their studies and their implications are described in detail in the extended summary and are available in Bulletin no. 45.
Project 109-08 | 3R-Info Bulletin 45 |
| | Completion of a project (March 2011) |  | Development of an in vitro assay for screening of antischistosomal drugs
Prof. Jennifer Keiser, Swiss Tropical and Public Health Institute, University of Basle, Switzerland Infection by the parasite Schistosoma, which causes bilharziasis, can be treated in humans by targeting juvenile or adult Schistosoma parasites (in blood or organs respectively). At present, the parasites are obtained using mice or hamsters and the efficacy of potential treatments is tested in mice that have been infected with the parasite. Unlike the adult stages, the juvenile form can be cultured in vitro. The studies carried out by Prof. Keiser have shown that substances that were not effective against the juvenile stages of the parasite also showed no effect on the adult stages. Through improved in vitro testing it will no longer be necessary to carry out subsequent tests on laboratory animals.
Project 110-08 |
| | Completion of a project (March 2011) |  | Reducing the number of fish and their suffering during acute toxicity testing of potential environmental pollutants (OECD Guideline no. 203).
Dr. Hans Rufli, ecotoxsolutions, Basle, Switzerland Through this project it has been possible to provide practical suggestions as to how the fish test involved in ecotoxicological studies (OECD guideline no. 203) can be improved from a 3R point of view. Thanks to retrospective analyses of hundreds of data series from fish tests and a mathematical simulation it has been possible to demonstrate that the number of fish used per test group could be reduced by 14% without any loss of quality in the results. A further reduction in the number of fish required can be achieved if the fish embryo test is used to determine the initial dose. The results obtained by Dr. Rufli’s research team have been discussed and accepted by selected experts in Europe and the USA. Since fish are caused level 3 suffering if the required maximum dose is used (and is effective), an attempt is being made to instigate a request from an OECD country (e.g. Switzerland) for a change in the OECD guidelines worldwide. The results of this project are summarised in Bulletin no. 43
Project 114-08 | 3R-Info Bulletin 43 |
| | Serum-free defined media, a largely unsolved problem in cell culture (February 2011) |  | 3R-Info-Bulletin 45 Cell culture methods enable the number of animal experiments in biological and industrial research to be reduced and in some cases replaced totally. Fetal calf serum must be added to most media to achieve optimum growth and to maintain cell functions. The composition of this serum is not defined, however, and varies from charge to charge. For reasons of animal protection, it would be preferable if the serum did not have to be taken from unborn calves. A partial step in this direction could be the identification of macro-molecular factors in the lipid protein-free fraction in fetal calf serum, as described in the Bulletin.
3R-Info Bulletin 45 | Project 109-08 |
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Deadlines Principal areas for financial support Instructions for applicants Application Form (Word) Form for yearly reports (Word) Guidelines for Awarding Research Grants Addresses 
European Consensus Platform for 3R Alternatives to Animal Experimentation
UK National Centre for the Replacement, Refinement and Reduction of Animals in Research
The global clearinghouse for information on alternatives to animal testing
Non-animal Methods for Toxicity Testing
The European Partnership for Alternative Approaches to Animal Testing
14.-16. March 2012-02-02 Technopark Zürich / Switzerland.
1st European Conference on the Replacement, Reduction and Refinement of Animal Experiments in Ecotoxicology
28th - 29th of June 2012. Eawag, Dübendorf, Switzerland.
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