Experts

Stichting Proefdiervrij: Collaboration is key
Expert interviews
HealthInnovationPolicy

Stichting Proefdiervrij: Collaboration is key

At Stichting Proefdiervrij (the Dutch society for the replacement of animal testing) we believe that collaboration is essential for the development and implementation of animal-free models. In this video we introduce a few of the ways in which we, as an NGO, collaborate with researchers to reach our goal: the complete replacement of all test on animals
02:0720 months ago
Monique Janssens (personal account): Why we need the Transition towards Animal-free Innovations
Expert interviews
HelpathonsPolicy

Monique Janssens (personal account): Why we need the Transition towards Animal-free Innovations

Why is there a Transition towards Animal-free Innovations, while we have the 3Rs, including Replacement? Well, there is a difference. Animal experiments should no longer be the golden standard of reference. We should not ask: Is this animal-free method good enough to replace animal experiments? But: What is the research question, and how do I get the best answer, preferably without animals? I know that many researchers are doing this already. But we can do even more! It’s also about involving the full chain of parties, including patients, financers, legislators and companies. That is why the transition movement works with interdisciplinary networks and Helpathons. The transition helps to innovate, to accelerate and to implement. At the same time, there is no need to throw the 3Rs overboard. Actually, we owe applying them to the lab animals of today. But by innovating we can develop even more new practices in research and education that bring about better results for science in less time and often with less costs. Without using animals.
02:372 years ago
Daniela Salvatori: TPI Utrecht
Expert interviews
HealthEducation

Daniela Salvatori: TPI Utrecht

Prof. dr. Daniela Salvatori, chair of TPI Utrecht, presents the aims of the local TPI group and invites all who want to share their ideas or questions on the transition towards animal-free innovations to get in touch via uu.nl/tpi.
02:233 years ago
Elly Hol (UMC Utrecht): possibilities for neuroscience
Expert interviews
HealthInnovation

Elly Hol (UMC Utrecht): possibilities for neuroscience

Prof. dr. Elly Hol (neuroscientist) talks about the opportunities for conducting animal-free research in Utrecht. She explains why it is necessary to use animal models next to cell-based models, for example for her Alzheimer research.
00:593 years ago

Innovation

New approaches for cancer hazard assessment
Innovation examples

New approaches for cancer hazard assessment

Chemical substances are subjected to assessment of genotoxic and carcinogenic effects before being marketed to protect man and the environment from health risks. For cancer hazard assessment, the long-term rodent carcinogenicity study is the current mainstay for the detection of nongenotoxic carcinogens. However, carcinogenicity studies are shown to have prominent weaknesses and are subject to ethical and scientific debate. A transition toward a mechanism-based weight of evidence approach is considered a requirement to enhance the prediction of carcinogenic potential for chemicals. At RIVM, we are working on this alternative approach for cancer hazard assessment, which makes optimal use of innovative (computational) tools and be less animal demanding. For more information, click on the link in the video or read on here (https://doi.org/10.1080/10408444.2020.1841732) and here (https://doi.org/10.1080/10408444.2018.1458818). Contact the expert (https://nl.linkedin.com/in/mirjamluijten)
03:142 months ago
Development of 3D liver spheroids
Innovation examples
HealthToxicologyInnovationIn vitro

Development of 3D liver spheroids

Human-based in vitro models are increasingly being used in the hepatology field. And in addition to the obvious ethical arguments, they offer several advantages over the classical animal models. One of them is the ability to perform mechanistic research at the molecular level in a well-controlled setting and reduce species differences. These liver-based in vitro models can range from simple monolayer cultures of hepatocytes to the liver-on-chips systems in which all liver cells are cultured in a 3D configuration on a microfluidic platform. Liver-based in vitro models must be selected on a case-by-case basis and should fit the purpose of the research, which might go from fundamental to translational research.
01:0410 months ago
Platform for in vitro airborne inhalation testing
Innovation examples
HealthToxicologyInnovationIn vitro

Platform for in vitro airborne inhalation testing

The air-liquid interface (ALI) technique uses lung cells cultured on a tiny polymer membrane in a cup. On one side of the membrane is a liquid containing the medium necessary for the cells to survive, while the other side is in contact with air. This is similar to the situation in the human lung. The compound to be tested is administered via an aerosol, vapor, or gas to mimic the situation in human lungs. By monitoring different parameters in the cell model before and after the compound is added, it is possible to measure the effects on lung cells. Depending on the test to be carried out, the lung cells can come from different regions in the respiratory tract and even from a variety of people, including individuals who smoke a lot or have specific diseases such as chronic obstructive pulmonary disease or asthma. In vitro ALI inhalation testing (https://doi.org/10.1021/acs.est.7b00493) adds value for e.g. pre-clinical trials and research in the pharmaceutical industry and testing (new) compounds for the chemical sector and beyond. The advantages of ALI inhalation testing are that it is a non-animal method, it reduces the use of in vivo experiments, pre-clinical testing with human-derived cell models is more realistic and limits clinical trial failures and it provides faster and more efficient testing of compound
04:1310 months ago
Using skin and mucosa models to replace animal testing
Innovation examples
HealthInnovationIn vitro

Using skin and mucosa models to replace animal testing

The skin and mucosa are important tissues that differ between species in health and disease. The group of Sue Gibbs works on the development of advanced in vitro models that mimic these two tissues, specialising in immunity models and organ-on-a-chip technologies. They use skin models to study for example melanoma, skin allergies, eczema, burns and healing wounds. Dental models are used for the safety of materials used in dentistry, for example to test the quality of the implant and false tooth when it comes to attaching to the soft tissue. Their ambition is to expand into the field of multi-organ technology to make even more relevant models for the human skin and mucosa. Click on the link in the video to watch more or read the interview with Sue he[https://vu.nl/en/research/more-about/using-skin-and-mucosa-models-to-replace-animal-testing]re.
00:3013 months ago

Meetings

Helpathon #10 – Can you help Jolanda and Elza?
Meeting videos
HelpathonsEducation

Helpathon #10 – Can you help Jolanda and Elza?

Jolanda van der Velden, Chair of Physiology, and Elza van Deel, Educator, from Amsterdam University Medical Center want to support PhDs in preparing for the animal-free transition. They are both looking for an implementation strategy and course design. Do you have an interest in animal-free education and education about animal-free research? Read more and register here (https://www.helpathonhotel.org/coming-up).
00:552 months ago
ONTOX Hackathon: Hack To Save Lives And Avoid Animal Suffering
Meeting videos
HelpathonsHealthToxicologyData

ONTOX Hackathon: Hack To Save Lives And Avoid Animal Suffering

Artificial Intelligence (AI) in toxicology – a potential driver for reducing or replacing laboratory animals in the future. ONTOX project is looking for solutions and innovative ideas to move forward. Are you going to help ONTOX to hack into these complex challenges? The hackathon will be held from 21 to 23 April 2024 in Utrecht Science Park. The whole event is open to a diverse community of forward-thinkers and problem-solvers interested in the intersection of AI and ethical toxicology. The goal is to bring together passionate individuals who seek innovative solutions to critical challenges in toxicology. Read more about the hackathon and register here (https://ontox-project.eu/hackathon/).
01:042 months ago
Helpathon #9 – Can you help Juan?
Meeting videos
HealthInnovationIn vitro

Helpathon #9 – Can you help Juan?

Juan is an experienced immunologist and scientific director of the cutting edge O2Flow facility for cytometry and cell sorting at the Amsterdam VU University Medical Center. Can you help Juan explore if and how he can transition his facility towards animal free antibodies? Are you using antibodies in your research do you want others to help you find animal free alternatives for your specific research let us know. More information can be found [here] (https://www.helpathonhotel.org/coming-up).
02:029 months ago
 Helpathon #8 – Can you help Margot?
Meeting videos
HealthInnovationIn vitro

Helpathon #8 – Can you help Margot?

Margot Beukers is the LymphChip program manager. Can you help Margot bring the field forward by sharing your experience with animal-free alternatives for Foetal Calf Serum and Matrigel? Click on the link in the video to sign up and read more information on this Helpathon on the website (https://www.helpathonhotel.org/coming-up).
01:0313 months ago

Projects and initiatives

Cells4Thought: using iPSCs for neurodevelopmental health
Projects and initiatives
HealthToxicologyInnovationIn vitro

Cells4Thought: using iPSCs for neurodevelopmental health

The prevalence of neurodevelopmental disorders (NDDs), including cognitive impairments, is increasing worldwide with great impact on daily life quality. There is evidence that exposure to chemicals may contribute to the incidence of NDD. However, a causal link is lacking. Towards this goal, a human-relevant in vitro model system mimicking parts of brain development, such as neuronal network functioning, could be used for mechanistic research on how gene-environment interactions contribute to the development of NDD. This is going to be studied in the project Cells4Thought, using induced pluripotent stem cells form different individuals to study the effect of chemicals on neuronal differentiation.
02:3824 days ago
We all want a safer world for humanity, animals and the environment: Transition Animal-free Innovation
Projects and initiatives
HealthInnovationPolicy

We all want a safer world for humanity, animals and the environment: Transition Animal-free Innovation

Why is the transition to animal-free research so important? What are animal-free models? How does TPI (Transition Animal-Free Innovation) encourage their development and use? And who are we working with to make this happen? We explain this in our animation. More and more animal-free tests and research methods are becoming available, but not all research questions or safety tests can be answered in this way yet. In addition, the validation, qualification and acceptance of non-animal innovations still lags behind. Therefore, the Dutch Ministry of Agriculture, Nature and Food Quality (LNV) stimulates the development and application of animal-free innovations. This is done with the partner programme Transition Animal-free Innovation (TPI).
02:482 months ago
VitalTissue: scientific research can be more human(e)
Projects and initiatives
HealthInnovationIn vitro

VitalTissue: scientific research can be more human(e)

The goal of VitalTissue is to facilitate the availability of vital human residual tissue for all researchers in the Netherlands. This video shows how VitalTissue works. From a request from a researcher, the donation of the residual tissue by the patient and the transport to the lab. This process is the result of a feasibility study conducted with many stakeholders. The national tissue bank ETB-BISLIFE will implement VitalTissue in practice.
04:218 months ago
In3: combining in vitro and in silico for chemical safety assessment
Projects and initiatives

In3: combining in vitro and in silico for chemical safety assessment

The in3 project aims to drive the synergistic development and utilisation of in vitro and in silico tools for human chemical and nanomaterial safety assessment. The project focused on differentiation of human induced pluripotent stem cells to toxicologically relevant target tissues including; brain, lung, liver and kidney. The tissues, from the same genetic backgrounds, were exposed to common compounds and the data generated and prediction tools generated were used to develop modernised safety assessment approaches combining cheminformatics, mechanistic toxicology and biokinetics into computational models which can account for donor and tissue specific effects.
03:2519 months ago

Conferences

Scientific solutions for the gap in translational medicine: skin model platform with melanoma (3D melanoma)
Conferences abstracts

Scientific solutions for the gap in translational medicine: skin model platform with melanoma (3D melanoma)

The developing process of a new drug, from first testing to regulatory approval and ultimately to market is a long, costly, and risky path. Noteworthy is the fact that almost 95% of the drugs that go into human trials fail. According to the National Institutes of Health (NIH), 80 to 90% of drug research projects fail before they ever get tested in humans. The value of preclinical research, mainly conducted in animal model experiments for predicting the effectiveness of therapies and treatment strategies in human trials, has remained controversial. Only 6% of the animal studies are successfully translated into the human response. Breaking down failure rates by therapeutic area, oncology disorders account for 30% of all failures. The absence of human-relevant models with receptors, proteins, and drug interactions in the in situ microenvironment leaves a gap in the scientific discovery process of new therapies. In this context, the present work presents the development of a sophisticated in vitro skin model platform focus on boosting melanoma treatment. The results showed a physiological microenvironment of human skin with epidermal differentiation and development of stratified layers (basement membrane, stratum spinosum, stratum granulosum, and stratum corneum). Furthermore, it was observed the pathophysiological microenvironment of the melanoma with invasion or migration through the basement membrane into the dermis and no epidermal differentiation. Vemurafenib treatment, the gold standard which targets BRAF mutations, showed a decrease in proliferation and invasion of melanoma tumors, with an increase in epidermis keratinization. Melanoma incidence continues to increase year-on-year and is currently responsible for >80% of skin cancer deaths. It is the most common cutaneous form and is known to have the highest mutational load of all cancers. Nowadays, patients with advanced melanoma BRAFV600E mutation can benefit from monotherapies or targeted therapies. Although the initial response rate is effective, disease progression and tumor chemoresistance rapidly occur in the majority of patients. Therefore, the treatment of melanoma remains a challenge, and despite the advances, there is still an urgent need to identify new therapeutic strategies. 3D Model Melanoma is considered one important tool for studying the evolution of the pathology, as well as evaluating the effectiveness of new therapeutic approaches.
03:192 years ago
Optimizing CAR-T-cell therapy using 3D tumor models and real-time cell imaging
Conferences abstracts

Optimizing CAR-T-cell therapy using 3D tumor models and real-time cell imaging

Chimeric antigen receptor (CAR) T-cell therapy accounts for one of the most promising therapeutic advances in cancer immunotherapy. In this form of adoptive cell transfer, T-cells of a patient are engineered to express so-called ‘CARs’, in which the antigen-recognition capacity of antibodies is combined with T-cell activating domains. So far, CAR-T-cell therapy obtained its most impressive results in hematological malignancies resulting in the approval of five CAR-T cell products by the FDA for hematologic indications. However, CAR-T-cell therapy has not mirrored its success in solid tumors. The poor efficacy of CAR-T-cell therapy in solid tumors has, in part, been attributed to the lack of understanding in how CAR-T-cells function in a solid tumor microenvironment. Classical validation methods rely on the use of specificity and functionality assays in 2D models against adherent target cells or target cells in suspension. Yet, by using these models, observations made in vitro may differ greatly to an in vivo situation where tumors are engrafted in 3D structures. We developed a more relevant and translational 3D tumor model using eGFP+ target cells. This allows us to couple 3D tumor cell killing by CAR-T-cells to live-cell imaging, providing an efficient quantification of target cell death. As proof- of-concept, we used a 3D model of eGFP+ glioblastoma cells and CAR-T-cells targeting a pan-cancer antigen. This 3D glioblastoma model allowed us to show that classical scFv-based CAR-T-cell therapy of glioblastoma cells can be improved by nanoCAR-T-cells. Furthermore, combining nanoCAR-T-cell therapy with a genetic approach of nanobody-based anti-PD-L1 immune checkpoint blockade further increased the cytotoxicity of the nanoCAR-T-cell therapy.
03:252 years ago
Biotransformation of two proteratogenic anti-epileptics in the zebrafish (Danio rerio) embryo
Conferences abstracts

Biotransformation of two proteratogenic anti-epileptics in the zebrafish (Danio rerio) embryo

The zebrafish (Danio rerio) embryo has gained interest as an alternative model for developmental toxicity testing, which still mainly relies on in vivo mammalian models (e.g., rat, rabbit). However, cytochrome P450 (CYP)-mediated drug metabolism, which is critical for the bioactivation of several proteratogens, is still under debate for this model. Therefore, we investigated the potential capacity of zebrafish embryos/larvae to bioactivate two known mammalian proteratogens, carbamazepine (CBZ) and phenytoin (PHE) into their mammalian active metabolites, carbamazepine-10,11-epoxide (E-CBZ) and 5-(4-hydroxyphenyl)-5-phenylhydantoin (HPPH), respectively. Zebrafish embryos were exposed to three concentrations (31.25, 85, and 250 μM) of CBZ and PHE from 51⁄4 to 120 hours post fertilization (hpf) at 28.5°C under a 14/10 hour light/dark cycle. For species comparison, also adult zebrafish, rat, rabbit and human liver microsomes (200 μg/ml) were exposed to 100 μM of CBZ or PHE for 240 minutes at 28.5°C. Potential formation of the mammalian metabolites was assessed in the embryo medium (48, 96, and 120 hpf); pooled (n=20) whole embryos/larvae extracts (24 and 120 hpf); and in the microsomal reaction mixtures (at 5 and 240 minutes) by targeted investigation using a UPLC–Triple Quadrupole MS system with lamotrigine (0.39 μM) as internal standard. Our study showed that zebrafish embryos metabolize CBZ to E-CBZ, but only at the end of organogenesis (from 96 hpf onwards), and no biotransformation of PHE to HPPH occurred. In contrast, our in vitro drug metabolism assay showed that adult zebrafish metabolize both compounds into their active mammalian metabolites. However, significant differences in metabolic rate were observed among the investigated species. These results highlight the importance of including the zebrafish in the in vitro drug metabolism testing battery for accurate species selection in toxicity studies.
02:572 years ago
Lung tumor spheroids for onco-immunological research
Conferences abstracts

Lung tumor spheroids for onco-immunological research

Lung cancer thrives in a complex multicellular tumor microenvironment that impacts tumor growth, metastasis, response, and resistance to therapy. While orthotopic murine lung cancer models can partly recapitulate this complexity, they do not resonate with high-throughput immunotherapeutic drug screening assays. To address the current need for relevant and easy-to-use lung tumor models, we established a protocol for fully histo-compatible murine and human lung tumor spheroids, generated by co-culturing lung fibroblasts with tumor cells in ultra-low adherence 96-well plates. Moreover, we describe their application potential to study tumor-stroma organization, T-cell motility, and infiltration as well as distinct macrophage subsets’ behavior using confocal microscopy. Finally, we report on a 3D target specific T-cell killing assay that allows spatio-temporal assessment using live cell imaging and flow cytometry. This lung tumor spheroid platform can serve as a blueprint for other solid cancer types to comply with the need for straightforward onco-immunology assays.
03:092 years ago