Experts
Expert interviews
Transition beyond animal welfare
This video explains what the programme TPI (Transition Programme for Innovation without the use of animals) is about.
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
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.
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.
Innovation
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.
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
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.
Innovation examples
HealthInnovationData
Using data and computational modelling in biomedical research
Bioinformatics and systems biology hold great promise to translate the wealth of biological data into meaningful knowledge about human health and disease. The group of Bas Teusink helps biologists to deal with high throughput data, for example metabolomics (how cell metabolism works) and proteomics (how protein networks work) from patient material or cell cultures. This can help to better understand disease mechanisms and aid drug targeting or personalised medicine. In the future, combining data from different models (in vitro, in vivo and human data) could become a digital model of humans, or a “ digital twin”.
Click on the link in the video to watch more or read the interview with Bas (and Jaap Heringa) he[https://vu.nl/en/research/more-about/using-data-and-computational-modelling-in-biomedical-research]re.
Meetings
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).
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).
Meeting videos
HealthInnovationIn vitro
Helpathon #8 – Can you help Jasper?
Jasper Koning is doing research on skin diseases. He believes it must be possible to find an alternative to Foetal Calf Serum to grow immune cells. Can you help him find alternatives to Foetal Calf Serum so he can build human models animal free? Jasper is especially looking for researchers with practical experience in applying alternatives. He did some trials himself with mixed results.
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).
Meeting videos
HealthInnovationIn vitro
Helpathon #8 – Can you help Germaine?
Germaine Aalderink is investigating the uptake of lipids travelling from the gut into the lymphatic system and further explore the merits of this alternative drug intake strategy. Can you help Germaine make an intestinal and lymphatic model with an alternative for Matrigel that is animal-free? She wants to know what components are essential in each phase of intestinal development and is interested in both the positive and negative experiences of other researchers with the use of alternatives for 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).
Projects and initiatives
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.
Projects and initiatives
In3
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.
Projects and initiatives
The Beyond Animal Testing Index
The Beyond Animal Testing Index (BATI) was designed after the Access to Medicine Index with the aim to be a transparent, objective and independent benchmark that provide public research organisations and their stakeholders insight in what efforts and contributions they make in the transition to animal free innovation and to provide organisations incentive to learn from and inspire each other with regard to the implementation of research practices without the use of animals for the benefit of science.
Projects and initiatives
SCREENED
The European collaborative project SCREENED aims to develop three-dimensional (3D) cell-based in vitro tests to better characterize the effects of endocrine disruptors (EDs) on thyroid gland function. This method will overcome the limitations of existing tests, being more sensitive at low doses of exposure to chemicals, and enabling the prediction of their toxicity on human health in a sex-specific manner. The ambition of the SCREENED project is that these new 3D in vitro tests, as well as the increased knowledge about adverse reactions after exposure to EDs, will be used for regulatory purposes, ultimately to improve human health.
Conferences
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.
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.
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.
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.
Other
Various subjects
InnovationPolicy
TPI.tv: improving science through animal-free innovations and research
Introducing TPI.tv : a video platform by experts striving to improve science through animal-free innovations and research.
Various subjects
Five simple tricks for making your own video for TPI.tv
This video shows you how to make a video yourself. It's really not that difficult! See also https://tpi.tv/how-to-submit for additional information.
Various subjects
ToxicologyDataPolicy
A New Way to Evaluate Chemical Safety and Assess Risk
TOX21