IC-3Rs • Innovation Centre for 3R Alternatives

How does the Brussels Region support IC-3Rs?

The Brussels Capital Region is very supportive of IC-3Rs efforts to promote the 3Rs principle and non-animal methods. Bruxelles Environnement provides financial support for:

  • Minister of animal welfare Bernard Clerfayt

    Minister Clerfayt is in the Brussels region responsible for animal welfare. As consequence, he has taken several initiatives to reduce the use of laboratory animals. For example, he supports the in vitroprojects that are carried out in the department of toxicology at the Vrije Universiteit Brussel. This gives young researchers the opportunity to gain experience and expertise

    in the development and use of human stem cell cultures to study the effects of drugs on the human liver. In addition, the Brussels government is actively supporting the RE-Place database, which is a joint project between Flanders and Brussels to centralize in a database all animal replacement methods for which expertise exists in Belgium. This gives young researchers easy access to these new technologies and allows them to gain experience in the laboratories to which they are affiliated.

  • Minister of animal welfare Bernard Clerfayt

    Le ministre Clerfayt est dans la région de Bruxelles responsable du bien-être des animaux. Donc, il a pris plusieurs initiatives pour réduire l'utilisation d'animaux de laboratoire. Par example, il soutient les projets in vitro qui sont menés dans le département de Toxicologie à la Vrije Universiteit Brussel. Les jeunes chercheurs ont ainsi l'occasion d'acquérir de l'expérience et de l'expertise

    dans le développement et l'utilisation de cultures de cellules souches humaines pour étudier les effets des médicaments sur le foie humain. En outre, le gouvernement bruxellois soutient activement la base de données RE-Place, qui est un projet conjoint entre la Flandre et Bruxelles, visant à centraliser dans une base de données toutes les méthodes de remplacement des animaux pour lesquelles il existe une expertise en Belgique. Les jeunes chercheurs ont ainsi un accès facile à ces nouvelles technologies et peuvent acquérir de l'expérience dans les laboratoires auxquels elles sont rattachées.

  • Minister of animal welfare Bernard Clerfayt

    Minister Clerfayt is verantwoordelijk voor het dierenwelzijn in het Brussels Gewest. Daarom heeft hij verschillende initiatieven genomen om het gebruik van proefdieren te verminderen. Zo steunt hij bijvoorbeeld de in vitro-projecten die worden uitgevoerd binnen de dienst Toxicologie van de Vrije Universiteit Brussel. Dat biedt jonge onderzoekers de kans om ervaring en expertise te verwerven

    in de ontwikkeling en het gebruik van menselijke stamcelculturen om de effecten van geneesmiddelen op de menselijke lever te bestuderen. Daarnaast ondersteunt de Brusselse regering uitdrukkelijk de databank RE-Place, een gezamenlijk project van Vlaanderen en Brussel om alle diervervangingsmethoden waarvoor in België expertise bestaat, in een databank te centraliseren. Op die manier krijgen jonge onderzoekers gemakkelijk toegang tot nieuwe technologieën en kunnen ze ervaring opbouwen in de laboratoria waaraan ze verbonden zijn.

Doctoral Theses

Running PhD thesis: Alexandra Gatzios

Partly sponsoring for 1 year: 2021: Development and application of a human stem cell-based in vitro  model for studying NASH-fibrosis

Metabolic-associated fatty liver disease (MAFLD) is one of the most prevalent chronic liver diseases worldwide. MAFLD encompasses multiple disease stadia, ranging from benign liver steatosis to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis and cancer. NASH often associates with liver fibrosis and is hence the tipping point to the life-threatening stages of MAFLD. Yet, no drugs against it are approved. This is partly due to a lack of suitable preclinical models. Animal models are not predictive and relevant for the human situation. Therefore, the prime objective of this research is to develop an in vitro human stem cell-based model that allows the investigation of NASH-related liver fibrosis. The project is supervised by Dr. Joost Bockmans.

Completed and partly sponsored PhD thesis: 2017-2020: Joost Boeckmans

Development and characterization of a human stem cell-based in vitro model for anti-NASH drug testing

Non-alcoholic steatohepatitis (NASH) is a severe chronic liver disease that affects about 5% of the population. NASH is characterized by hepatic lipid accumulation, inflammation and fibrosis and can progress to cirrhosis and hepatocellular carcinoma. There are currently no drugs available to treat NASH. Investigation of NASH traditionally relies on animal models, which are often not representative for the human situation. Therefore, the aim of the doctoral thesis was to develop a human-based in vitro model that can recapitulate the molecular and cellular mechanisms that drive NASH and can be used during anti-NASH drug development. The project was supervised by Prof. Robim M Rodrigues, Prof. Tamara Vanhaecke, Prof. Vera Rogiers and Prof. Joery De Kock from the Vrije Universiteit Brussel.

Completed and partly sponsored PhD thesis 2017-2020: Alessandra Natale

Shift from 2D to 3D in the development of a human skin cell-derived hepatic in vitro model for toxicological applications

Drug-induced liver injury (DILI) is a major threat to human health and is, together with absence of clinical efficiency, at the basis of a 90% failure rate in drug development. As preclinical data obtained using experimental animals do not adequately represent the human situation, in vitro models based on human cells are highly needed. In this doctoral thesis, the relevance of human-skin derived stem cells differentiated towards hepatic cells (hSKP-HPC) for toxicological screening was highlighted and the biological importance of ‘in vivo-like’ conditions for culturing and differentiating human stem cells was emphasized. Although the process requires further optimization, the results pave the way to enhance hepatic maturation which is a prerequisite to reliably predict human-specific hepatotoxicity of new pharmaceuticals. The project was supervised by Prof. Robim M Rodrigues, Prof. Tamara Vanhaecke, Prof. Vera Rogiers and Prof. Joery De Kock from the Vrije Universiteit Brussel.

in collaboration with Sciensano funded by the Departments Animal Welfare of the Flemish region (Department ‘Omgeving’) and Brussels region (former Kabinet Debaets, current cabinet Clerfayt)

The RE-Place database

RE-Place is a scientific project funded by the Flemish and Brussels government which aims to collect the available expertise on the use of alternative methods to animal testing, also referred to as ‘New Approach Methodologies’ (NAMs), in one central database.

This open access database is available for different stakeholders such as the scientific community, the government, regulators, ethical committees and other interested parties. It provides a reliable overview of the available knowledge on NAMs, but also the names of experts who you can contact and research centres where these techniques can be learned in Belgium.



In order to obtain such an up-to-date and reliable inventory, we need the help of the scientific community in Belgium. We invite you to submit your knowledge on the NAMs in which you have experience via the RE-Place online tool (www.RE-Place.be). These methods do not necessarily have to be developed by yourself, nor by the organization you are working for. Your personal know-how on NAMs is the essential information we are looking for.



By submitting your expertise, you and your organisation can demonstrate your commitment to animal-free innovation and you have many different advantages:

  • Increasing the visibility of your work to other scientists,
  • Facilitating the search to be identified as an expert in the field,
  • Exchanging experiences on the use of NAMs, learning from others and fostering new collaborations,
  • Allowing the identification of knowledge gaps which will help to better allocate future funding,
  • And so much more!



We are collecting information on all NAMs in basic and applied research that avoid the direct use of animals, used as stand-alone or combined with animal experimentation to collect the information of interest. NAMs are thus not necessarily one-by-one replacement methods but can contribute to the overall replacement or reduction of animal testing. They can also be a single step within a broader research strategy.

NAMs thus include different types of:

  • In vitro and ex vivo methods (e.g. experiments with the use of 2D - 3D cell lines and tissue cultures, NRU Phototoxicity Test, AMES, BCOP, …);
  • In silico modeling (e.g. molecular modeling and mathematical approaches, PBPK models, QSAR, read across, …);
  • In chemico techniques (e.g. assays evaluating the reactivity and properties of substances or components);
  • Alternative in vivo models (e.g. fruit flies, flatworms, early stages of zebrafish, …);
  • Other innovative techniques (e.g. organ-on-a-chip);



The RE-Place project is coordinated by the Vrije Universiteit Brussel and Sciensano. More information via the website www.RE-Place.be and on our YouTube Channel.

Do not hesitate to contact info@RE-Place.be or mieke.vanmulders@sciensano.be if you have questions.


Completed doctoral theses
  • 2017-2020: PhD Joost Boeckmans
  • 2017-2020: PhD Alessandra Natale
Running doctoral theses
  • Starting October 2018: Quinten Marcelis
  • Starting October 2019: Anouck Thienpont
  • Starting February 2021: Alexandra Gatzios
Running doctoral theses
  • Starting October 2018: Quinten Marcelis
  • Starting October 2019: Anouck Thienpont
  • Starting February 2021: Alexandra Gatzios

Current status of the RE-Place database comprising expertise on the use of NAMs in Belgium

RE-Place is a scientific project of the Flemish and Brussels Capital regions which aims to centralize the available knowledge on the use of alternative methods to animal testing, also referred to as ‘New Approach Methodologies’ (NAMs), in Belgium. The collected information is integrated in an open access database which can be freely accessed by the scientific community, the government, regulators, ethical committees and other interested parties. The RE-Place database not only provides a reliable overview of the available expertise on NAMS, but also the names of experts who you can contact and research centres where these techniques can be learned. By facilitating access to the available knowledge and improving communication, RE-Place will grow as a powerful tool to stimulate the use of NAMs and promote their (further) development.

The ‘News and Events’ section of the RE-Place website provides information on the latest developments and events happening in the field of NAMs.

Vision on the use of test animals

Vision on the Use of Test Animals


The medical scientific mission  of the Vrije Universiteit Brussel (VUB) entails that, to fulfill its social responsibility, it conducts innovative research aimed at preventing, predicting and curing diseases, among other things.

In doing so, the VUB strives for an optimal research environment by providing the necessary support for researchers and by enabling and requiring continuous training. Moreover, every VUB researcher signs the "charter of the good researcher" that includes clear guidelines on correct scientific-ethical behaviour as well as the regulations on scientific integrity violations. All this means that research is carried out in a strictly regulated framework. When conducting scientific research at the VUB, not only legal obligations are examined, but also social-ethical considerations. Thus, every researcher must ask himself the question "What is the most appropriate model to answer my research question?" beforehand. Examples of appropriate models are a computer simulation, performing tests on cells and tissues of laboratory animals and/or humans (in vitro research) or performing experiments on laboratory animals (in vivo research). The VUB is strongly committed to the development of non-animal test methods through the expansion of the IC-3R Centre. The  Innovation Centre for 3R Alternatives (IC-3Rs) was founded at the VUB on 25 September 2017. This platform aims to stimulate the development, visibility and use of alternative 3R methods (Replacement, Reduction, Refinement) - and to strengthen the communication on this subject, building on the extensive expertise of the IVTD (In VitroToxicology and Dermato- Cosmetology) group. Furthermore stimulate the development of in vitro methods by building on the long-standing activities and expertise of the IVTD research group.  Growth is possible thanks to the  VUB Chair/legate Mireille Aerens for the development of non-animal test methods and the support of the  Brussels Region (Minister Bernard Clerfayt). The focus is on giving opportunities to young researchers and innovative projects to conduct research in this field and thus broaden the basis for non-animal research. IC-3Rs aims to use fewer animals wherever scientifically possible and to increase the focus on integrating alternative non-animal methods into basic and applied research, the areas where most animals are used worldwide.

"Can we develop new medicines without animals today?" All experts clearly agree: today this is not yet possible, but we can carry out parts of the research without using animals among others by using sophisticated cultures of human stem cells...Should animal testing be necessary within a research project, those involved must adhere to the legal and ethical standards set out in (inter)national laws and regulations.  These state, among other things, that any project which requires the use of laboratory animals must receive ethical approval from a legally recognised Experimental Ethics Committee (ECD) before it can begin.An important question which the Ethical Committee on Animal Experiments of the VUB always asks itself during the evaluation of a research project is: "What is the social and scientific added value of the expected results of the research and does this added value outweigh the discomfort the test animal might experience?".

If the research on animals has an important added value, we will always strive to apply the legal principles of the 3Rs as much as possible. On the one hand, this means that researchers always use the lowest possible animal species and keep the number of animals to a scientifically justified minimum. On the other hand, the animals must be housed in the best possible conditions and their welfare must be monitored by the researchers, the veterinarians experts and the members of the Animal Welfare Unit. Today, the "best science" is a well-considered combination of in vivo and in vitro methodology, and the follow-up of new developments is crucial so that non-animal techniques can be integrated as soon as they are well established and available. With the construction of a new VUB animalarium, considerable investments have been made in recent years that allow for further optimised housing and animal care. The 3Rs are further complemented by the VUB with the principles of "Commitment" and "Accountability". Every researcher has the obligation to follow training and education and every researcher has to give account to both official bodies and the general public. One of the ways in which the VUB is replacing or reducing the use of laboratory animals is by promoting the use of  RE-Place among researchers. This database, supported by Animal Welfare Flanders and the Brussels Region, provides a reliable overview of "New Approach Methodologies (NAMs)", which reduce or even eliminate the need for laboratory animals. In addition, the tool can be used to identify experts and research centres where the techniques can be taught..

At the VUB, there are also several European research projects that endorse the 3Rs, such as Ontox, Twinalt, PARC... 

Furthermore, the VUB signed  the transparency agreement on animal testing in Belgium. The aim of this Agreement is to ensure that members of the Belgian public receive accurate and up-to-date information on animal testing:

  • how such research is regulated in Belgium
  • the role it plays in the overall process of scientific discovery, treatment development and safety testing
  • the efforts made by researchers and staff to support animal care and welfare
  • what is being done to reduce the use of animals and minimise their suffering.