Charité 3^R Toolbox

Definition

The replacement of animal experiments with methods or testing strategies not entailing the use of living animals.
(source: German Centre for the Protection of Laboratory Animals, Bf3R)
Further definitions of the 3Rs can be found on the NC3Rs website.

LAB CONSUMABLES

NC3Rs animal-free in vitro technologies
This online resource helps in vitro researchers to move to animal-free options with confidence, providing all the information needed to make the switch for more reproducible and reliable experiments. The resource covers alternatives to animal-derived antibodies, culture media and scaffolds, sharing the benefits of animal-free technologies, case studies, lists of suppliers and more.

Animal-free cell culture media
Fetal Calf Serum (FCS, also known as Fetal Bovine Serum, or FBS) is a common supplement of animal cell culture media. Provided by the 3Rs-Centre ULS in collaboration with Animal Free Research UK, this database provides an overview of FCS-free media for cell-culture.

The EURL ECVAM Recommendation on Non-Animal-Derived Antibodies
This science-for-policy report describes the EURL ECVAM recommendations on non-animal-derived antibodies developed on the basis of the ESAC Opinion (Annex 1) and its detailed Working Group Report (Annex 2). The review focused on non-animal-derived antibodies generated by phage-display technology. FAQs on this recommendation are available, too.

Biocompare's database for Recombinant Antibodies
Recombinant Antibodies are produced without the use of laboratory animals. Researchers can use the Biocompare database to search for them. Biocompare is a resource for up-to-date product information, product reviews, and new technologies for life scientists.

HUMAN TISSUE

Central Biobank Charité
This biobank is as a core facility for use by Charité and BIH researchers. Through the web-based project inquiry portal, interested scientists and external partners can apply for the use of biomaterials and data for research purposes and arrange the storage of future samples.

The German Biobank Registry
The German Biobank Registry provides an overview of all medically relevant biobanks in Germany. In addition to contact details for each of the registered biobanks, it also provides information on specific research foci and types of samples held.

German Biobank Node / German Biobank Alliance
German Biobank Node serves as a central cooperation platform for the German biobank community. The ‘Sample Locator’ tool enables users to search the biobanks which form part of the German Biobank Alliance (GBA) for samples with specific criteria. The German Biobank Directory provides key information on academic biobanks in Germany.

LITERATURE SEARCH

The EURL ECVAM search guide: Good search practice on animal alternatives 
This Guide was developed by the European Union Reference Laboratory for alternatives to animal testing (EURL ECVAM) to support untrained database users in finding high quality information on relevant alternative strategies and methods from the wide range of sources available on the web.

ALTBIB – Alternatives to Animal Testing
This NIEHS database on alternatives to animal testing provides a survey of the literature in a format which facilitates easy scanning. The bibliography includes citations from published articles, books, book chapters, and technical reports.

Alternatives search
This guide from the UC Davis Library explains the entire process of an alternatives search: how to define the right questions, select appropriate databases, construct a search strategy, evaluate search results, and manage search results.

3R Smart: Literature searches for alternatives to animal experiments (Presentation with audio file)
Using the example of the toxicity test for botulinum toxin, Dr. Barbara Grune (Bf3R) explains the search strategy for alternative methods. She is using PubMed and Livivo in this example.

DATABASES

EURL ECVAM Database on Alternative Methods to Animal Experimentation (DB-ALM) 
DB-ALM focuses primarily on methods submitted to EURL ECVAM for validation, and those identified in ad hoc reviews of the literature and of end-users in specific application areas between 2000 and 2019.

Alternative methods for toxicity testing: validated test methods 
The European Union Reference Laboratory for alternatives to animal testing (EURL ECVAM) forms an integral part of the Joint Research Centre (JRC), the European Commission’s science and knowledge service. It has contributed to the validation of the test methods listed here.

Definition

Reduction in the number of laboratory animals used to a minimum.
(source: German Centre for the Protection of Laboratory Animals, Bf3R)
Further definitions of the 3Rs can be found on the NC3Rs website.

Charité FEM animal pool
The FEM animal pool is still in the test phase and is only accessible to Charité scientists (via the Charité intranet). The aim of the animal pool is to transfer animals which cannot be used in planned animal experiments to other Charité research groups.

SYstematic Review Centre for Laboratory animal Experimentation (SYRCLE)
SYRCLE is the research and education center of the Animal Research Facility (CDL) of the Radboud University Medical Center in the Netherlands. A review article published by SYRCLE and entitled ‘A step-by-step guide to systematically identify all relevant animal studies’, asserts that “before starting a new animal experiment, thorough analysis of previously performed experiments is essential from a scientific as well as from an ethical point of view.”

Interspecies Database
Anatomical, physiological and biochemical differences are the main factors causing discrepancies between humans and animals with respect to the kinetics of xenobiotics., Launched by the National Institute of Public Health and the Environment (RIVM) in the Netherlands and 3Rs-Centre Utrecht Life Sciences (ULS), this database enables researchers to choose the best research model. Users can choose from different species and strains, organs, age classes, gender and specified parameters and compares these with each other or with human data. 

AniMatch: Sharing animal resources
AniMatch is an online platform which enables scientists to connect and share the organs and tissues of test animals. After registration, AniMatch allows scientists to post and search offers for the multiple use of test animals. This inspires responsible use of a valuable biological resource and reduces animal usage effectively.

Models in Translational Oncology (MiTO)
The MiTO platform which is run by the German Cancer Research Center (DKFZ), forms a central part of the German Cancer Consortium. The platform allows scientists to enter their cancer models and browse a wide spectrum of other existing models including animal models, patient samples, genetically homogenous permanently growing cell lines, and genetically heterogeneous primary cells derived from tissues of human patients or animals.

SEARCHBreast
The SEARCHBreast platform, which was established through NC3Rs funding, is a large network of breast cancer researchers with in vivo, in vitro, and in silico models of breast cancer. SEARCHBreast facilitates the sharing of archived material derived from in vivo breast cancer models.

Munich MIDY Pig Biobank
The Munich MIDY Pig Biobank facilitates systematic studies of organ crosstalk in diabetes in a multi-organ, multi-omics dimension. MIDY pigs represent a model of poorly controlled diabetes mellitus (DM) in humans. This complex biobank was built from 2-year-old MIDY and wild-type pigs.

QPS Austria GmbH - Rodent Biobank
QPS Austria is a full-service contract research organization (CRO) which focuses on CNS diseases, orphan diseases and mental disorders, and provides preclinical as well as clinical research services. Tissues of transgenic and non-transgenic animals of various age groups are readily available, such as brain tissue, CSF and plasma samples. The biobank is composed of various specimens derived from their in-house in vivo models.

Definition

Reducing the suffering of animals used in research
(source: German Centre for the Protection of Laboratory Animals, Bf3R)
Further definitions of the 3Rs can be found on the NC3Rs website.

Charité-internal Refinement-Sharepoint
This Charité-internal platform offers information on current refinement methods from selected external resources as well as the referring experts at Charité - Universitätsmedizin Berlin. Dear Charité colleagues, take the chance to get in contact and share your refinement experiences! (access restricted to Charité-affiliates)

Norecopa`s Refinement Wiki
This Wiki provides a platform to publish methods for the refinement of animal experiments. Find useful protocols and share your Refinement protocols with the research community.

Pain assessment in rodents
‘Facial action units’ increase in intensity in response to post-procedural pain and could therefore be used as part of a pain assessment. The UK National Centre for the Replacement, Refinement & Reduction of Animals in Research (NC3Rs) provides different grimace scales for mice, rats and rabbits.

Pain management in laboratory rodents (presentation and audio file)
Dr. Pauline Jirkof (Swiss 3R Competence Center) presents currently available methods for pain management in laboratory rodents and discusses their advantages and disadvantages.

Humane endpoints
This website from the 3Rs-Centre Utrecht Life Sciences (ULS) helps people to refine their experiments, improve their knowledge of animal welfare and enhance the quality of their research. It provides information about humane endpoints and teaches users how to recognize humane endpoints in rodents and fish.

NC3Rs Rodent Welfare Hub
NC3Rs provides information on a variety of topics, such as how to pick up a mouse; genetically altered mice; housing and husbandry. Please also use the search mask on the new structured NC3Rs resource library.

Animal Welfare Information Center
The Animal Welfare Information Center (AWIC) is an institution of the National Agricultural Library of the U.S. Department of Agriculture (USDA). AWIC provides comprehensive information pertaining to the fields of laboratory animal science and animal husbandry.

Critical incident reporting system in laboratory animal science (CIRS-LAS)
This critical incident reporting system is being established by ann interdisciplinary group of researchers from the University Hospital Jena. Incidents can be reported by anyone and are always treated confidentially. CIRS-LAS has reached productive state and has received registrations from several European countries (Germany, Austria and Switzerland). Incident reports will be published in the member area after review.

RSPCA – How to reduce severe suffering in lab animals
The Royal Society for the Prevention of Cruelty to Animals (RSPCA) is a registered U.K. charity which is dedicated to animal welfare. The RSPCA's primary aim is the replacement of animal experiments with humane alternatives worldwide. The RSPCA works with scientific institutions to provide information on the severe suffering of lab animals and how to reduce it.

Definition

Animal research often lacks measures to reduce validity threats such as biases or a lack of statistical power. Measures to reduce such bias, like randomization and blinding, are essential to ensure robust research results alongside sufficient numbers of experimental units.
(source: BIH QUEST Center)

Experimental Design and Statistical Analysis 
Section 4, ‘Experimental Design and Statistical Analysis‘, of the PREPARE guidelines provides a lot of information about this topic, including links to online protocols and further reading.

Experimental Design Assistant (EDA)
NC3Rs Experimental Design Assistant helps to design robust study protocols which are more likely to yield reliable and reproducible results. Protocols can be time-stamped and made publicly available (optional) to facilitate pre-registration.
EDA also provides information regarding components and concepts which are  crucial to experimental design, e.g. group and sample size, blinding, allocation.

Responsible PrecliniX
Responsible PrecliniX offers support concerning the model (including potential alternatives), statistics (sample size calculation), experimental design, robustness, and the role or need of replication studies (within a laboratory or between laboratories). Scientists can approach the team early on at the stage of proposal writing and Responsible PrecliniX accompanies projects from the basic idea to translation through a series of Go/No Go decision points. 

Service Unit Biometry
The Service Unit Biometry (Institute of Biometry and Clinical Epidemiology, Charité) provides statistical consulting services for scientists working at Charité and the BIH and helps researchers to put their research projects on a biometrically valid basis.

Mouse Models – White Papers and Handbooks
Commercial suppliers of laboratory animals also provide helpful handbooks and white papers:
Charles River – Strategies to Minimize Genetic Drift and Maximize Experimental Reproducibility in Mouse Research
The Jackson Laboratory - Handbook on Genetically Standardized Mice (Sixth Edition)
Taconic – White Papers

Definition

Publication bias is prevalent in animal research, meaning that null and negative results often end up in the file drawer, unpublished. Studies should be pre-registered (e.g. on https://www.animalstudyregistry.org/) to reduce publication bias and limit researcher degrees of freedom in analysis and tested hypotheses.
(source: BIH QUEST Center)

Animal Study Registry 
The Animal Study Registry is run by the ‘German Centre for the Protection of Laboratory Animals’ (Bf3R) at the German Federal Institute for Risk Assessment (BfR). The registry is intended to improve research quality and animal welfare (Heinl et al. 2020). It provides a platform to register an exact study plan for studies involving animals prior to the start of experiments. Each study receives a DOI (Digital Object Identifier) number - which records intellectual property ownership. Study visibility can be restricted for a period of up to five years.

Preclinical trials
This register is provided with the support of the Transnational AllianCe for regenerative Therapies In Cardiovascular Syndromes (TACTICS), but the register is open for studies from all fields of the biomedical sciences. The register aims to promote the online registration of preclinical trials to prevent publication bias, and replicates arrangements already in place for clinical trials (Jansen of Lorkeers et al. 2014).

Open Science Framework (OSF) Preregistration 
OSF Preregistration is part of the activities of the Center for Open Science, a non-profit organization. They stress, that it is important to separate hypothesis-generating (exploratory) from hypothesis-testing (confirmatory) research, see.

AsPredicted
The website AsPredicted is not a registry but a simple form of pre-registration that requires only what is necessary to separate exploratory from confirmatory analyses. However, researchers can post completed forms to a registry to meet preservation and transparency standards.

Definition

Published results often lack important information that is needed for a critical appraisal, such as information on study design and attrition of animals. Authors should therefore adhere to reporting guidelines such as ARRIVE and include all necessary information in their publication.
(source: BIH QUEST Center)

ARRIVE guidelines – Animal Research: Reporting of In Vivo Experiments 
The ARRIVE guidelines, originally published in PLOS Biology, were developed in consultation with the scientific community as part of an NC3Rs initiative to improve the standard of reporting of research using animals. The revised ARRIVE guidelines 2.0 are available in different languages, including German.

Equator network – Search for reporting guidelines 
The Equator network (Enhancing the QUAlity and Transparency Of health Research) provides a database with more than 400 reporting guidelines for all kinds of study types. The Equator Wizzard  helps authors find the correct reporting guidelines for their studies.

Review Article: Be positive about negatives – recommendations for the publication of negative (or null) results
This encouraging review article (Bespalov et al. 2019) describes a set of criteria that can help researchers publish technically sound, scientifically high-impact negative (or null) results originating from rigorously designed and executed studies.

BIH QUEST: Fiddle – file drawer data liberation effort
Fiddle is a tool developed by QUEST to combat publication bias. This ‘match-making’ tool helps researchers to identify alternative ways of publishing information from well-designed experiments which is often difficult to publish in traditional journals (i.e. null or neutral results, datasets, etc.). These alternatives to traditional journals include data repositories, micropublication, preprint publication, data journals, publishing platforms, and journals open to null results.

Charité Animal Welfare Officers’ Link List / Linkliste der Tierschutzbeauftragten der Charité
Prepared by Charité’s Animal Welfare Officers, this link list provides links to the following: relevant legal provisions; information on animal welfare and the 3Rs (including supplementary and alternative methods);  experimental animal science; and information centers.

NC3Rs resource hubs
NC3Rs is the UK's National Centre for the Replacement, Refinement & Reduction of Animals in Research. This comprehensive NC3Rs resource site includes several information hubs, such as e-learning resources, a rodent welfare hub, and an experimental design hub.

Norecopa
Norecopa is Norway's National Consensus Platform for the advancement of ’the 3 Rs’. Norecopa provides an International Meetings Calendar and Newsfeed, and issues an electronic newsletter 7-8 times a year. Norecopa published the PREPARE: Guidelines for planning animal research and testing.

The QUEST Toolbox
The BIH QUEST Center for Transforming Biomedical Research aims to increase the value and impact of biomedical research by maximizing its quality, reproducibility, generalizability, and validity. Their toolbox is focused on the reproducibility of animal experiments. It contains helpful tools, programs and online platforms – non-commercial wherever possible – and is organized according to the stages in the life cycle of a research project: preparation, execution and publication.

3Rs INFO HUB
The 3Rs INFO HUB pooles the most-relevant and latest approaches in 3Rs to one learning platform. Instead of getting lost in thousands of search engine results, the information is readily available. Courses and interactive QUIZZES consolidate and apply what has been learned. The PODCAST series "StudentCast on 3Rs" offers the latest discussion of students with international experts on the story behind alternatives to animal testing.

3R-SMART – Alternativmethoden für Tierversuche
3R-SMART (3R-Schulungsplattform für Methodische Ansätze zur Reduktion von Tierversuchen) is a new training platform for methodological approaches to reducing animal experiments. Contributions on this page are available in German and/or English. Most of these contributions are audio recordings of lectures accompanied by the corresponding presentations. A BMBF-funded project, the platform is coordinated and developed by the University of Veterinary Medicine Hannover Foundation and the Philipps Universität Marburg.

European Commission – Animals used for scientific purposes
All animal studies have to be carried out in compliance with EU legislation. Since 1986, the EU has had in place specific legislation covering the use of animals for scientific purposes. In 2010 the EU adopted Directive 2010/63/EU on the protection of animals used for scientific purposes. It took full effect on 1 January 2013.

Links to German animal welfare legislation (German only website)
The Charité animal welfare officers provide a comprehensive list of links to applicable animal welfare legislation and other relevant information.

PREPARE
PREPARE stands for ’Planning Research and Experimental Procedures on Animals: Recommendations for Excellence’. A group of experts from the UK and Norway, led by the Secretary of Norecopa, has produced a set of guidelines for planning animal experiments. They complement reporting guidelines such as ARRIVE and cover all stages of quality assurance, from the management of an animal facility or population to the individual procedures which form part of a study.

ARRIVE
ARRIVE stands for ’Animal Research: Reporting of In Vivo Experiments‘. Originally published in 2010, the ARRIVE guidelines were developed as part of an NC3Rs initiative to improve standards of reporting and ensure that the data from animal experiments can be fully evaluated and utilized. In 2020, the guidelines have been updated and information reorganized to facilitate their use in practice (find here original articles on the ARRIVE guidelines 2.0 and explanations on the updates).

GIVIMP 
GIVIMP stands for ’Guidance Document on Good In Vitro Method Practices‘. It is part of the OECD Series on Testing and Assessment  and addresses the development and implementation of in vitro methods for regulatory use in human safety assessments. It provides guidance on key elements of in vitro methods for test method developers and the end users of research data.

Guideline on the principles of regulatory acceptance of 3Rs testing approaches
This EMA guideline defines regulatory acceptance and gives guidance on the scientific and technical criteria for regulatory acceptance of 3Rs testing approaches. Pathways for regulatory acceptance of 3Rs testing approaches and procedures for submission and evaluation of a proposal for regulatory acceptance of 3Rs testing approaches are described.

FAIR 
The FAIR Guiding Principles for scientific data management and stewardship state that data must be Findable, Accessible, Interoperable, and Reusable (Wilkinson et al. 2016). These guidelines put specific emphasis on enhancing the ability of machines to automatically find and use the data, in addition to supporting its reuse by individuals.