General Outcome - COST Action IMPROVE

On the 13th of June 2026 at the R3FINE conference at the Medipol University in Istanbul, Turkiye, Winfried Neuhaus was invited to give a talk on "Animal-free drug testing –a vision or already reality ?". He provided an overview about the implementation of NAMs into the drug development pipeline, the current developments in the 3Rs and NAMs field including several policy initiatives and the COST Action IMPROVE.

Abstract: Animal-free drug testing is increasingly transitioning from an aspirational concept to an emerging component of regulatory science and pharmaceutical development. This presentation examines the current scientific and regulatory status of New Approach Methodologies (NAMs) within the broader framework of the 3R principles – Replacement, Reduction, and Refinement – and evaluates whether a paradigm shift is already underway. Recent advances in human-relevant in vitro systems, including organoids, microphysiological systems (organ-on-chip platforms), induced pluripotent stem cell technologies, and advanced 3D tissue models, have enabled the generation of mechanistically informed and human-centered data. These approaches are increasingly combined with computational modeling, physiologically based pharmacokinetic simulations, artificial intelligence, and omics technologies to improve predictivity and translational relevance. Regulatory integration of NAMs is being accelerated through international initiatives such as OECD Integrated Approaches to Testing and Assessment (IATA), activities coordinated by ICCVAM and the FDA in the United States, and validation efforts led by EURL ECVAM in Europe.
Particular attention is given to the role of European 3R centers coordinated through EU3Rnet, which facilitate knowledge exchange, training, and strategic alignment across member states. In addition, collaborative frameworks such as the COST Action IMPROVE and COST Action TEATIME are strengthening interdisciplinary cooperation, promoting standardization, and advancing translational implementation of NAMs.
Although challenges remain in validation, regulatory acceptance, and systemic integration, substantial progress has been achieved. Animal-free drug testing can therefore no longer be regarded merely as a vision, but rather as a developing reality within defined contexts of use.

The main congress homepage could be found via following link: r3fine.org

The abstract band could be found via following link: R3FINE_2026.pdf

The main program could be found via following link: R3FINE-090626-program.pdf

Fernando Gonzalez-Uarquin, Paulin Jirkof, Bettina Bert, Penny Hawkins, Ljupco Angelovski, Jan Baumgart, Nadine Baumgart, Özge S Cevik, Nuno H Franco, Erdal Horata, Rohish Kaura, Winfried Neuhaus, Brigida Riso, Adrian J Smith, Athanassia Sotiropoulos, Augusto Vitale, Sophie Schober. Building bridges: Involvement of animal care staff and laboratory technicians in experimental planning and conduct of animal studies for better job satisfaction and science. Lab Anim (2026). doi: 10.1177/00236772251400976.

The involvement of non-scientific staff in discussions about animal welfare and scientific quality is essential for biomedical research progress. In this study, we developed a survey to collect the self-perception of animal care staff (ACS) and laboratory technicians about their involvement in scientific planning and conduct. Participants were contacted to complete an anonymous online questionnaire. We obtained 850 responses, mainly from Europe: 564 from ACS and 286 from laboratory technicians. Job satisfaction was assessed as positive by ACS and laboratory technicians despite the low frequency of culture of care activities and mental health meetings. Both groups expressed their desire to be trained in research planning and conduct; however, regular training was not reported. In addition, the inability to act on animal welfare concerns owing to experimental reasons was reported by both groups. Over half of the participants felt valued and appreciated by the lead scientists or animal facility manager; however, it is not clear how they are acknowledged, as their names on the authors list or in the manuscript acknowledgments are barely included. Our results indicated that involvement of ACS and laboratory technicians in planning and conducting studies would improve their understanding of how experiments are done, and therefore communication processes, work satisfaction, animal welfare, and scientific quality. Finally, we provided recommendations to improve the engagement of ACS and laboratory technicians in discussions about animal research planning and conduct.

Arti Ahluwalia, Babette Fletemeyer, Giuseppe Chirico, Alisa Elezovic, Winfried Neuhaus, Adrian J Smith, Jeffrey J Bajramovic, Doris Wilflingeder. What's in a NAM ? Lab Anim (NY) (2026). doi: 10.1038/s41684-026-01731-8.

The transition away from animal experimentation is gaining momentum across scientific disciplines, driven by technological advancements, ethical imperatives and policy support. At the forefront of this movement is the development of New Approach Methodologies (NAMs). Rather than serving as mere substitutes for animal testing, NAMs represent a paradigm shift. Their growing acceptance has been reinforced by recent policy advancements, such as the US Food and Drug Administration Modernization Act 2.0 and the European Chemicals Agency’s commitment to transitioning toward an animal-free regulatory system, both of which promote the adoption of NAMs. However, the term NAMs has become increasingly ambiguous, leading to confusion and potential misuse. To address this, we propose a unified definition: NAMs are species-specific methodologies, not including the use of living animals. NAMs prioritize the use of the target species in some form or another, eliminating the need for interspecies extrapolation. If the target species is human, this simple definition implies that working with experimental animals as a model never qualifies as a NAM. This distinction positions NAMs as a fundamentally different approach to the 3Rs framework of Replacement, Reduction and Refinement, and practical examples are discussed. Critically, this definition extends beyond toxicology and regulatory science, encompassing basic, applied and translational research. By leveraging cutting-edge technologies, including organoids, organ-on-chip, microfluidics, 3D bioprinting, bioinformatics, artificial intelligence and machine learning, NAMs provide novel biological insights that differ from those obtained through animal models while enhancing species-relevant data accuracy. Although challenges remain, particularly in their validation and widespread adoption, continued innovation, policy support and interdisciplinary collaboration will be key to unlocking their full potential, fostering more sustainable, humane and human-relevant approaches in biomedical research.

On the 30th of April 2026 at the Austrian 3Rs Days Winfried Neuhaus was invited to give a talk on "An overview of international and European developments in the 3Rs and NAMs field". He provided an overview about the current developments in the 3Rs and NAMs field including several policy initiatives and the COST Action IMPROVE.

Abstract: The transition toward animal-free biomedical research and safety assessment is gaining momentum worldwide, driven by scientific innovation, evolving regulations, and policies promoting the 3Rs: Replacement, Reduction, and Refinement. This presentation highlights recent international and European developments in New Approach Methodologies (NAMs), focusing on policy initiatives and collaborative networks that support non-animal approaches. Internationally, major policy and funding efforts are accelerating the integration of NAMs into biomedical research and regulatory science. In the United States, recent versions of the FDA Modernization Act
and strategic initiatives by the National Institutes of Health (NIH) support the use of human-relevant in vitro and in silico methods in drug development and safety testing.
In Europe, the policy landscape is evolving rapidly. European Commission activities, including work on a roadmap to phase out animal testing for chemical safety assessments, reflect a strong strategic commitment to NAMs. Collaborative frameworks such as EPAA and EU3Rnet foster cooperation among regulators, industry, and academia. New EC and Horizon Europe initiatives, including the European Research Area on NAMs and the project NAMWISE,
further promote regulatory uptake through guidance, training, and stakeholder engagement. Together, these efforts mark substantial progress in integrating NAMs into research and regulatory practice.

The main congress homepage could be found via following link: Highlights 2026 - Austrian 3R-Days

The abstract band could be found via following link: Abstractband-3Rdays-2026.pdf

The main program could be found via following link: Hauptprogramm-3Rdays-2026-Final.pdf

Céline Heinl, Nuno H Franco, Bettina Bert, Katherina Siewert, Dania Movia, Edlira Aruçi, Carina Ladeira, Ahmed Y Sanin, Winfried Neuhaus, Ulf D Kahlert, Manosij Ghosh, Aurélie Thomas & Athanassia Sotiropoulos. A call for preregistration of in vitro research: Pregistration as a tool to improve reproducibility and transparency. EMBO Reports (2026). doi: 10.1038/s44319-026-00764-x.

In this episode, Prof. Alistair Lawrence from Roslin Institute of the University of Edinburgh, Scotland, presented about "What can rats teach us about positive animal welfare".

This video is based upon work from COST Action AFFECT-EVO (CA23106), IMPROVE (CA21139) and LIFT (CA21124), supported by COST (European Cooperation in Science and Technology). COST is a funding agency for research and innovation networks. COST Actions help connect research initiatives across Europe and enable scientists to grow their ideas by sharing them with their peers. This boosts their research, career and innovation; www.cost.eu.

A series of webinars organized by the IMPROVE, AFFECT-EVO and LIFT COST Actions, dedicated to improving animal welfare in research and promoting the principles of the 3Rs (Replacement, Reduction, Refinement). These sessions feature leading experts discussing best practices, innovative approaches, and ethical considerations in laboratory animal science.

In this episode, Dr. Laura Webb from Wageningen University, the Netherlands, presented about "animal happiness - understanding and assessment through behaviour, cognition and physiology". She is co-chair of the COST Action LIFT.

📍 Porto, Portugal | 🗓️ 8–10 October 2025

The IMPROVE COST Action (CA21139) successfully hosted its first Training School at i3S – Instituto de Investigação e Inovação em Saúde, bringing together educators, researchers, and trainers from across Europe to explore innovative approaches to teaching the 3Rs (Replacement, Reduction, Refinement) in animal research.

Key Outcomes:

Pedagogical Innovation: Participants engaged in hands-on sessions focused on education science, learning how to effectively teach the 3Rs using modern, learner-centered methods.
Ethical & Emotional Dimensions: The program emphasized the importance of addressing emotional and ethical aspects in laboratory animal science education.
Collaborative Exchange: The hybrid format enabled broad participation, fostering international collaboration and knowledge sharing.

Impact

This Training School marked a significant step in building a community of educators committed to improving research quality and animal welfare through education. It laid the groundwork for future initiatives and collaborations within the IMPROVE network.

🔗 For more details and resources, visit: https://cost-improve.eu/outcomes/

Booklet_-3RS_In-person Download

Genova, 23^rd September 2025 - The COST Action IMPROVE successfully held its next 3Rs centres Workshop at the Università degli Studi di Genova, Dipartimento Architettura e Design (DAD), gathering leading researchers and stakeholders from across Europe.

Over two days from 17^th to 18^th of September, participants engaged in collaborative sessions focused on the 3Rs (Replacement, Reduction, and Refinement) and the role and future activities within the 3Rs centre network EU3Rnet.

The workshop opened with remarks from Arti Ahluwalia and Winfried Neuhaus, followed by updates on the Memorandum of Understanding and the Balkan Regional 3R Centre.

Attendees took part in discussions on communication, funding, and sustainability, and enjoyed valuable networking opportunities, including a group dinner in the heart of Genova. The second day featured sessions on the organisation and management of EU3Rnet, short pitches on emerging topics, and the formation of new working groups to address key challenges in the field.

The hybrid format allowed both in-person (about 40) and online participation, ensuring broad engagement and collaboration.

This event was held together with the Next Gen3Ration Research meeting of the Italian 3R centre (Centro 3R; https://www.centro3r.it/it/events/next-gen3ration-research) from 18^th to 19^th of September.

A heartfelt thank you to the organisers of this remarkable event - Elena Grasselli and her team !

In this episode, Prof. Georgia Mason presents about proxy measures for animal feelings: how to validate indicators of affective state or cumulative affective experience.

Vitale Augusto, Simone Pollo, Nikos Kostomitsopoulos, Aurelie Thomas, Gamze Kaya, Kaan Bicici, Yesim Isil Ulman. The Istanbul Workshop organised by the ‘IMPROVE’ COST Action Ethics Crossover Group. Altern Lab Anim (2025). doi: 10.1177/02611929251345074.

The overall aim of the ‘Improving the Quality of Biomedical Science with 3Rs Concepts’ (IMPROVE) COST Action (CA21139) is to establish an interdisciplinary network dedicated to refining, harmonising and promoting the adoption of the Three Rs concepts, to enhance the quality of biomedical research. The ‘IMPROVE’ COST Action has assembled an Ethics Crossover Group composed of volunteer members from each of IMPROVE’s four established Working Groups: ‘Quality and translatability of science’, ‘Implementation’, ‘Dissemination’ and ‘Education’. This report relates to a two-day workshop, held in Acibadem University in Istanbul on 2–3 September 2024. The workshop was organised by the Ethics Crossover Group in line with the aims and the scope of the ‘IMPROVE’ COST Action, to address the ethical dimension of research and experiments on laboratory animals.

Through this STSM, I aimed at gaining epertise in working with human intestinal organoids, to further enrich the Home Institution's expertise in developing more physiological relevant biomimetic in vitro intestinal models.

With regard to the accomplished practical work, initially, human-derived intestinal organoids were maintained in culture for 7 days (Figure 1) and subsequently passaged or differentiated into apical-out configuration (Figure 2), allowing me to become familiar with key aspects of their handling, differentiation, and maintenance.

Figure 1. Human jejunal organoids in culture for 7 days (Magnification: 4x).

Figure 2. Differentiated human jejunal organoids into apical-out configuration (Magnification: 40x). Another major goal of this STSM was to evaluate the intestinal permeability of exenatide-based delivery systems using this 3D intestinal model. Permeability studies were conducted by testing 1 µM of exenatide released from delivery systems, using human intestinal organoids differentiated into the apical-out configuration, after 24 hours.

In parallel, I acquired hands-on training in the development of monolayers derived from intestinal organoids—another key objective of this STSM. Organoids were enzymatically dissociated into single cells and seeded with different cells densities onto inserts (24-well plate format) coated with a Matrigel/Collagen I mixture (1:1). Media was changed every other day for 5 days and TEER values were measured to ensure the formation of a complete monolayer (Figure 3).

Figure 3. Monolayers-derived from human intestinal organoids seeded at different cell densities.

As an alternative approach to evaluate the efficacy of the delivery systems in the absence of human pancreatic islets, a bioluminescence resonance energy transfer (BRET)-based assay was employed to monitor the activation of the GLP-1 receptor (GLP-1R). This method enabled us to assess the functional engagement of the receptor by the released recombinant exenatide, providing valuable insights into the bioactivity of the delivery systems.

In summary, a new collaboration was established between the Home (i3S, Porto, Portugal) and Host Institution by my STSM focusing on the development of advanced in vitro intestinal models to be used as an alternative to animal models for drug testing. This transfer of knowledge contributes to the reduction of animal experiments used for drug testing and nanomedicines, in accordance with the goals of working group 1 of the CA IMPROVE. I have received specific training in culture of human intestinal organoids and in the establishment of monolayers-derived form intestinal organoids that will be applied at my Home Institution to improve the reproducibility and physiological relevance of the in vitro intestinal model.

Beyond scientific outcomes, the STSM contributed significantly to my personal and professional development. The opportunity to work in a new research environment abroad enriched my doctoral training, in line with the goals of working group 4 of the CA IMPROVE.

Maria Kitsara, presented a poster at FELASA 2025 entitled "IMP3ROVE Animal Research" under the Topic 5 - Aiming for a better future, highlighting the impactful work COST Action IMPROVE - 3Rs concepts to improve the quality of biomedical science (CA21139). This initiative is dedicated to advancing the 3Rs principles—Replacement, Reduction, and Refinement— to elevate the quality and ethical standards of biomedical science.

The presentation showcased:

the mission and activities of IMPROVE
the current status of the network in terms of participants
the key outcomes achieved so far,

This work underscores the importance of responsible research practices and the power of international collaboration in shaping the future of biomedical science

At the session titled “Do I have a choice? Reconciling personal and professional ethics in laboratory animal science training” Augusto Vitale presented a talk called “I am not sure I want to do this…: students’ emotional perspective on animal experiments”.

In this presentation he presented qualitative results form the interviews with 16 students at the istituto Superiore4 di Sanità in Rome. The interviews focused on the potential emotional difficulties encountered with experimental animals, and the way to solve such difficulties. The self-trust in carrying out such procedures in a professional manner was the main factor tu overcome the unpleasentness of the procedures. The sacrifice of animals resulted to be the most critical aspect. The majority of respondants can do it, when they manage to put aside the emotional aspect of killing a living being, and put forward the need for science. The ones who cannot follow this mental process will not sacrifice animals. Related to this, it appeared that one-on-one relationship with a tutor or supervisor was by far preferred to the participation in organised courses.

The crucial factors in addressing by staff members emotional distress in students were: give time and respect; never force anybody to do anything they dont feel like doing; understanding emotional diversity; empathy and dissipation of fears; re-locate tasks within a project.

Augusto Vitale and Anna Olsson jointly organized a session entitled “Do I have a choice? Reconciling personal and professional ethics in laboratory animal science training”.

Session summary:

The use of animals in research is ethically problematic, in particular through the harm that is caused to experimental animals. Much has been invested in addressing the ethical issues on a collective level (e.g. regulations, training, licensing and ethics review).

Much less attention has been given to the moral quandary of animal experimentation in the context of individual ethics. It has been left to each (future) professional in the field (graduate students, researchers, animal technicians, veterinarians) to find their own way of dealing with the ethical and emotional challenges implied in a profession where harming and killing animals is sometimes unavoidable. This can no longer be taken for granted. As teachers and mentors, we increasingly meet early career researchers in training who find themselves in a real quandary as regards their own use of animals in experiments. We argue that it is time to discuss how to best support these future professionals. We believe that the typical research community responses to trainees’ concerns (claiming that they like everyone else need to get used to the practice, or questioning if they are a good fit for the job) are problematic. We will argue that as established practitioners in the laboratory animal science community, we can play a key role in supporting a more constructive discussion about when to use animals and how to use them, hence ultimately contributing to reducing animal suffering.

This session is organized in collaboration with COST Action CA21139 3Rs concepts to improve the quality of biomedical science (IMPROVE)

Session program:

Sonja Rumpel (Germany): Psychological Stress and Strain in Laboratory Animal Professionals

Renelle McGlacken (United Kingdom): Personal Impact of Animal Use: (Tangential) Insights from an interview study with UK researchers/students

Augusto Vitale (Italy): I Am Not Sure I Want to Do It: Emotional Perspective on Animal Experiment

Anna Olsson (Portugal): Who Should Learn to Do Experiments with Animals and How?

At the session titled “A Win-Win Situation for Research Quality and Work Satisfaction: More Involvement of Animal Care Staff” (organised by Fernando Gonzales Uarquin and Paulin Jirkof) Augusto Vitale presented a paper called “The role of technicians in qualitative assessment of animal welfare and the subjective experience”.

In this paper he addressed the role of technicians in the qualitative assessment of animal welfare, particularly in the context of non-human primate research. The presentation underscored that while objective welfare indicators are important, technicians—through their daily interactions and familiarity with individual animals—can provide valuable insights into psychological wellbeing and individual differences. Empathy and mutual trust between staff and animals were highlighted as important factors in welfare assessment. Empathy was also identified as a common and necessary requisite to carry out daily maintenance of animal colonies.

At the FELASA 2025 Congress in Athens, the SkillUp Tech session titled “A Win-Win Situation for Research Quality and Work Satisfaction: More Involvement of Animal Care Staff” brought together professionals from across Europe to discuss the evolving role of animal care staff and technicians in laboratory animal science. The session, chaired by Fernando Gonzalez Uarquin and Paulin Jirkof, focused on the potential benefits and current challenges of increasing the involvement of these staff members in research planning and conduct. A quick on-site survey revealed that a high number of lab technicians and animal care staff participated in the session.

Survey Insights: The current situation of Communication and Involvement

Sophie Schober presented findings from a large-scale, European survey conducted under the COST Action IMPROVE initiative. The survey, which gathered responses from more than 30 countries, revealed several key points:

Overall, researchers rated communication with other project members as good, but actual discussion about experimental planning with animal care staff occurred infrequently.
Animal care staff and technicians expressed a clear interest in being more involved in discussions about planning and conducting experiments, as well as in receiving training on experimental planning and protocol development.
Both researchers and non-scientific staff recognized the value of such involvement for improving research quality

The Importance of Multidirectional Communication

Bettina Bert’s presentation highlighted that while animal experiments are essential for scientific progress, questionable research practices—such as poor planning, selective reporting, and lack of transparency—can undermine both the scientific and ethical value of these studies. Open science practices like preregistration, data sharing, and comprehensive reporting have the potential to address these issues, but successful implementation depends on all participants, including technicians and care staff, understanding the importance of research rigor and transparency. Bettina also presented results from the survey, ensuring the willingness of animal care staff to be involved in scientific discussions and indicating that there is still room to enhance communication and training in these areas together with animal care staff and lab technicians.

Qualitative Assessment of Animal Welfare

Augusto Vitale’s talk addressed the role of technicians in the qualitative assessment of animal welfare, particularly in the context of non-human primate research. The presentation underscored that while objective welfare indicators are important, technicians—through their daily interactions and familiarity with individual animals—can provide valuable insights into psychological wellbeing and individual differences. Empathy and mutual trust between staff and animals were highlighted as important factors in welfare assessment. Empathy was also identified as a common and necessary requisite to carry out daily maintenance of animal colonies

Interactive Discussion and Polling

The session included interactive polls, inviting participants to reflect on their professional background, experiences with compassion fatigue, and views on the involvement of care staff and technicians in experimental planning. The responses indicated a general openness to increased inclusion, particularly when supported by appropriate training and communication structures. It was also discussed that publishers could contribute to this process Lab technicians and animal care staff should be named in the acknowledgement or they should get the possibility to publish their opinions.

S. Schober, B. Bert and A. Vitale at the session

The poster dealt with the issues of communication, inclusion, and research culture within laboratory animal science supporting the central message of the session "SkillUp Tech session titled “A Win-Win Situation for Research Quality and Work Satisfaction: More Involvement of Animal Care Staff" involving all staff who contribute to animal research—especially those responsible for daily animal care—can lead to improvements in both animal welfare and research quality. Therefore, attention must be paid to training needs and implementation.

The day featured a dedicated workshop organized by BIOEMTECH, centered on the practical application of the 3Rs principles within a preclinical CRO setting, specifically in the field of radiopharmaceuticals. With 34 participants attending on-site, the workshop showcased how BIOEMTECH embeds ethical excellence into every aspect of its operations.

The day opened with a series of presentations from the BIOEMTECH team, covering:

the scientific and technological scope of the company’s work
the fundamentals of radiopharmaceutical use in preclinical research
practical strategies for implementing the 3Rs — not only as theoretical guidelines, but as daily operational standards.

The agenda included the following talks:

Welcome address
Dr. Maria Kitsara, Science and Innovation Manager at BIOEMTECH
Introductory talk on BIOEMTECH’ journey
Dr. George Loudos, CEO, BIOEMTECH
BIOEMTECH CRO Lab - Diving into preclinical research for drug discovery
Dr. Maritina Rouchota, Chief Business Officer, BIOEMTECH
Animal models used in theranostics
Dr. Theodoros Karampelas, CSO, BIOEMTECH Laboratories
Animal welfare when working with radiopharmaceuticals & virtual tour in BIOEMTECH facilities
Argyro Zacharioudaki, Designated Veterinarian & Laboratory Animal Facilities Manager

During the afternoon, 20 participants had the opportunity to visit BIOEMTECH’s imaging systems manufacturing facilities & the CRO.
The lab visits contained:

Visit at the in vivo imaging systems manufacturing facilities
Visit at the preclinical CRO facilities - Demonstration in practice

1. Quality assurance of radiochemical compounds for animal administration

2. How to handle radioactive animals

3. Intravenous administration of radioactive compounds

4. Imaging with BIOEMTECH eyes.

From advanced imaging systems that allow short, non-invasive scanning — reducing stress and eliminating the need for more invasive procedures — to a dedicated CRO team that upholds animal welfare throughout the research process, the 3Rs are deeply integrated into BIOEMTECH’s core mission.

In this episode, Dr Tom Smulders presents on: "Adult hippocampal neurogenesis as a valence marker". Tom Smulders is Chair of COST Action AFFECT-EVO https://affect-evo.eu/.

On the 20th of May 2025 at Norecopa's 18th Annual Meeting Winfried Neuhaus was invited to give a talk on "The rise of 3Rs centres in Europe and their role in the development of the field of 3Rs". He provided an overview about the rise of the 3Rs centres in the last decades linked to the implementation of EU Directive 2010/63 EU, the current developments in the 3Rs field including several policy initiatives and the COST Action IMPROVE.

On May 15, 2025, the Young Researchers and Innovators (YRI) and the mentorship committees from COST Action IMPROVE hosted the online webinar "Ethics, Animal Research and One Health: Fostering Awareness and Career Development through 3R Principles and Animal Welfare," moderated by Anil Can and given by Dr. Yesim Isil Ulman. The event brought together approximately 50 participants from diverse backgrounds, including early-career researchers, animal welfare advocates, and professionals in biomedical sciences.

Event Highlights

The webinar provided a comprehensive overview of ethical considerations in animal research, with a particular focus on the 3R principles and their critical role in advancing both animal welfare and scientific integrity.
Participants engaged in discussions on the interactions of animal research ethics and the One Health approach, emphasizing the importance of responsible research practices for human, animal, and environmental health.
The session featured interactive elements, encouraging attendees to share experiences and reflect on challenges faced in their own research environments.

Key Outcomes

Increased Awareness: Attendees discussed the ethical frameworks guiding animal research and its relationships with the One Health concept.
Career Development: The webinar highlighted how ethical awareness and adherence to animal welfare standards can positively impact career progression in research, aligning with the broader goals of responsible science.
Networking and Inclusivity: The event fostered a supportive environment for knowledge exchange, connecting participants from various countries and disciplines, including those from Inclusiveness Target Countries (ITCs).

For further resources on ethics in animal research and career development, participants are encouraged to engage with the ongoing activities of COST Action IMPROVE (https://cost-improve.eu/).

During my Short-Term Scientific Mission (STSM) at the Toxalim DR2 INRAE Institute in Toulouse, France, I worked on non-genotoxic carcinogens (NGTxCs)—chemicals that induce cancer without directly damaging DNA. In addition to advancing my theoretical understanding of these substances, I gained practical expertise in developing and analyzing advanced in vitro 3D co-culture models. Throughout this project, I worked under the mentorship of Dr. Marc Audebert. My primary focus during the STSM was to establish and characterize a 3D co-culture system using three different liver cell lines: HepaRG (human liver cells), h-TERT-HSC (hepatic stellate cells), and TPH-1 (monocytic cells).

The aim was to create a more complex, physiologically relevant model to study NGTxC-induced toxicity. Using magnetic beads (NanoShuttle-Grainer), I integrated these three cell types. The magnetic beads facilitate cell aggregation, promoting the formation of spheroids that more closely mimic the structural and functional complexity of human liver tissue. These beads are biocompatible, composed of gold, iron oxide, and Poly-L-Lysine, and they do not affect cell proliferation, viability, metabolism, oxidative stress, or the phenotype. Furthermore, they do not interfere with experimental techniques such as immunohistological labeling, fluorescence detection, HR microscopy, Western Blot, or PCR/qPCR.

Once the multi-cellular co-culture system was established, I characterized it in terms of cell viability, differentiation, and interactions, ensuring it was suitable for toxicological testing. The INRAE institute is a global leader in developing systems that replicate human physiology, particularly in the fields of toxicology and biomedical research. Their expertise in tissue engineering and in vitro technologies proved invaluable throughout this process.

After learning how to maintain the co-culture system, I focused on applying confocal microscopy to analyze fixed 2D and 3D cell samples. I first learned how to fix and stain both monolayer cells and multicellular spheroids specifically for confocal imaging. Using advanced staining techniques, I labeled the spheroids with specific markers and fluorochromes to visualize cellular morphology, protein localization, and interactions within the co-culture system. The host institution’s extensive experience with confocal microscopy enabled high-resolution imaging of cellular interactions and structural changes, allowing for detailed analysis of these complex biological systems.

Once the co-culture models were established and characterized, I exposed the spheroids to selected NGTxCs and non-carcinogenic chemicals (as controls) to evaluate their effects. The exposure lasted for 24 and 96 hours (with repeat treatments) using five different compounds: etoposide (genotoxic), colchicine (genotoxic), KBrO3, D-mannitol, and a control. For each treatment, four spheroids were exposed to five concentrations of each compound in 10-fold dilutions. All experiments were performed in triplicate.

To assess the impact of the treatments, I first observed structural changes in the treated spheroids using confocal microscopy. This high-resolution imaging provided insights into cellular morphology, protein localization, and the interactions between cells within the co-culture system. I used a panel of five cell markers—DAPI (for nuclei), pH3 (for mitotic cells), GammaH2A.X (for double strand breaks), HMOX1 (for oxidative stress response), and IL-6 (for inflammatory response)—to characterize cellular responses to the treatments.

Additionally, we collected samples for further transcriptomic analysis, which will take place at the National Institute of Biology (NIB). This analysis will assess changes in gene expression triggered by NGTxCs, identifying the molecular pathways involved in NGTxC-induced carcinogenesis. These insights are critical for understanding the cellular mechanisms driving NGTxC toxicity.

The outcomes of the STSM directly support the objectives of the COST Action IMPROVE by:

Advancing scientific understanding of NGTxC mechanisms,
Contributing to the development and validation of alternative testing strategies (NAMs),
Strengthening international collaboration in predictive toxicology, and
Promoting knowledge transfer and capacity building between partner institutions.

In conclusion, the STSM successfully met its scientific, technical, and collaborative goals. It contributed meaningfully to the development of New Approach Methodologies (NAMs) and established a strong basis for long-term international cooperation under the framework of the COST Action IMPROVE.

Thriving during your career as a researcher? YES - and it can be learned!

On April 29, 2025, Dr. Felisa Berenguer, Training and Career Development Officer at ABG - Association Bernard Gregory, led the interactive online seminar “Successfully Navigating Your Research Career.” The event was organized by the YRI (Young researcher and innovators) and Mentorship Committees of the European COST Action IMPROVE, bringing together early career researchers from across disciplines and countries.

During the 1.5-hour session, Dr. Berenguer shared practical tools, actionable strategies, and insightful perspectives to help researchers maximize their professional potential while safeguarding their well-being. The seminar adopted a panoramic approach, addressing key aspects of a successful research journey:

Time Management & Planning Tools: Participants learned how to organize their projects efficiently, anticipate necessary actions, and plan ahead to meet deadlines without unnecessary stress.
Effective Communication Strategies: The session explored ways to communicate assertively, formulate clear requests, and navigate professional environments with confidence.
Recognizing Early Warning Signs: Dr. Berenguer highlighted the importance of self-awareness, teaching attendees how to detect early signs of stress or disengagement and take proactive steps to maintain mental health.

The webinar was designed to be highly interactive, encouraging participants to engage and reflect on their own experiences. The session’s inclusive format ensured accessibility for researchers from diverse backgrounds, including those from Inclusiveness Target Countries (ITCs).

Through this initiative, ABG - Association Bernard Gregory reaffirmed its long-standing commitment to supporting early career researchers with resources that help them care for themselves, pursue their ambitions, and build sustainable, fulfilling career paths.

Key Takeaways:

Research careers are marathons, not sprints - good project management, communication, and self-care are essential for long-term success.
Practical tools and strategies can help researchers navigate challenges and maintain well-being.
Professional development is a continuous journey, and support networks like ABG and COST IMPROVE are here to help every step of the way.

For more information about ABG’s career development resources, visit www.abg.asso.fr

Date: April 9–10, 2025

Location: University of Helsinki

Over two days, the FIN3R Symposium and Workshop brought together experts, researchers, and professionals from laboratory animal science to discuss and advance the culture of care in Finland and Europe. The event featured lectures from international and Finnish speakers on the first day, followed by an interactive workshop on the second day.

COST Action IMPROVE participated through Fernando Gonzalez Uarquin’ s presentation on integrating animal care staff and lab technicians into experimental planning—a promising step toward enhancing transparency in research practices. Such initiative, created within CA IMPROVE, involve around 14 members of the Action.

On Day 1, the symposium was chaired by Mikko Karpale (University of Oulu) with lecturers from Paulin Jirkof (University of Zurich, Switzerland), Fernando Gonzalez Uarquin (University Medical Center Mainz, Germany), Jordi Tremoleda (Queen Mary University of London, UK), Vootele Voikar (University of Helsinki), Hajnalka Nadai (Experimentica), and Hanna-Marja Voipio (University of Oulu). The discussion panel was chaired by Noora Holk (Orion Pharma), allowing participants to exchange ideas and reflect on the day's topics.

On day 2, the workshop focused on practical aspects of communication challenges within animal facilities. Led by Jordi Tremoleda and Fernando Gonzalez Uarquin, it brought together 15 participants for an interactive session.

Key activities included: 1) Sharing Care-full stories to illustrate real-world experiences, 2) Group discussions addressing routine communication problems encountered in animal facilities. 3) Exploring training materials designed to adapt culture-of-care practices to different institutions. 4) Exchanging ideas and experiences among participants from diverse professional backgrounds.

For further inquiries about the FIN3R Symposium and Workshop or to learn more about the culture of care initiatives, please reach out to the organizing team via email at info@fin3r.fi. Additional resources, including training materials and networking opportunities, can also be accessed through the FIN3R website: www.fin3r.fi.

The presentation of my research, "Immunological and Oxidative Stress Responses of N/TERT-1 and N/TERT-2G Keratinocyte Cell Lines," at the Society of Toxicology 64th Annual Meeting and ToxExpo significantly elevated my professional visibility and catalyzed the formation of valuable new contacts for future collaborations, particularly in the context of a Marie Skłodowska-Curie Actions (MSCA) Postdoctoral Fellowship grant proposal. During my poster session attracted considerable attention due to its focus on the translational relevance of N/TERT keratinocyte models for studying immunological and oxidative stress mechanisms in skin biology. My findings' scientific depth and practical applications resonated with academic and industry stakeholders.

These interactions not only broadened my professional network but also allowed me to assess the potential for assembling multidisciplinary industrial partners for the MSCA Postdoctoral Fellowship proposal. The exposure from my presentation, combined with proactive engagement during the conference, has positioned me as a key connector in opening the doors to innovative collaborations that bridge academic and industry expertise in cell biology, organ-on-chip technology, and toxicity assessment.

A photo standing beside my poster during the Late-breaking poster session.

The 4^th Asian Congress for Alternatives to Animal Experiments (4ACAAE) and 7^th Annual Meeting of the Society for Alternatives to Animal Experiments India (SAAE-I) took place in Jamia Hamdard, New Delhi, India from 12.12.-14.12.2024 (www.jamiahamdard.ac.in/4ACAAE) under the leadership of Prof. Dr. Y.J. Gupta and the Local Organizing Committee (LOC), led by Prof. Seikh Raisuddin (Organizing Secretary) supported by Prof. H.A. Khan, Prof. Suhel Parvez, Dr. Nidhi and many more wonderful people from his team.

Winfried Neuhaus was invited to the congress and gave a talk in the international session entitled “Overview about recent European 3Rs activities with a focus on 3Rs centres”, in which he took a closer look at current European developments in the 3Rs field with respect to the EC roadmap of phasing out animal studies and the response dynamics from several stakeholders, and the activities of EU3Rnet (network of European 3R centres) and the COST Action IMPROVE.

Introduction and Updates of the COST Action IMPROVE; Talk of W. Neuhaus at the SGV 2024 in Lausanne, Switzerland (26.-27.2024). Link to the event: SGV Meeting 2024

W.Neuhaus, réunion SGV 2024, 27 novembre, 2024, CHUV, Lausanne. (© Gabriel Monnet)

A report about the COST Action IMPROVE was issued on the website of the German 3Rs Network "Bundesnetzwerk-3R.de". The report can be found via following link:

https://www.bundesnetzwerk-3r.de/DE/3r-auf-einen-blick/themen/praxistipps/_documents/improve/improve.html?nn=1060654

In addition to the description of the COST Action project IMPROVE, Action chair Winfried Neuhaus answers three questions. The article is written in German language.

Introduction and Updates of the activities of EU3Rnet including an introduction of the COST Action IMPROVE; Talk of W. Neuhaus at the Animalfree Research Forum (Forum - Animalfree Research) in Zurich, Switzerland (11.11.2024).

A summary video of the event could be found via this link: Animalfree Research forum 2024

Kitsara M, Smajlhodžić-Deljo M, Gurbeta Pokvic L, et al.

"Introducing the COST Action ‘Improving the Quality of Biomedical Science with 3Rs Concepts’ (IMPROVE)."

Alternatives to Laboratory Animals. 2024;52(6):326-333. doi:10.1177/02611929241286024

The European Union-funded COST Action, ‘Improving the Quality of Biomedical Science with 3Rs Concepts’ (IMPROVE), which kicked-off in October 2022, presents an unprecedented opportunity to harmonise and synergise efforts in moving the Three Rs forward. European Cooperation in Science and Technology (COST) is a funding organisation for research and innovation networks, supporting interdisciplinary research networks to investigate a specific topic from all different research fields. The ‘IMPROVE’ COST Action aims to establish an interdisciplinary network dedicated to refining, harmonising and promoting the adoption of the Three Rs principles, in order to enhance the quality of biomedical research.

Lauwereyns, J., Bajramovic, J., Bert, B. et al. Toward a common interpretation of the 3Rs principles in animal research. Lab Anim (2024). https://doi.org/10.1038/s41684-024-01476-2

Despite their broad acceptance, interpretations of the 3Rs vary substantially, shaping statutory frameworks at various levels, with both technical and practical impacts.The work tries to contribute to a common understanding of the 3Rs.

During the three sessions, eight participants shared their experiences on how a growth environment can propel performance, resilience, creativity, and psychological safety—all essential for tackling complex biomedical science challenges. By fostering a culture of psychological safety and promoting learning agility, the course helped attendees cultivate a more open, creative, and solution-driven mindset, critical to advancing the 3Rs principles in biomedical research.

The attendants of the MTG program had access to the innovative Growth Indicator psychometric tool, which helped them to identify and challenge mental habits limiting their potential. This tool consists of a pretest before the course and a post-test six months after, allowing the measurement of long-term progress. In that way, the participants highlighted the course's accessibility, with long-term access to training materials (up to 12 months) allowing for sustained habit changes and continued personal development.

The feedback emphasized the program's positive impact on enhancing collaboration and fostering a nurturing research environment, aligning perfectly with the goals of the CA21139 - IMPROVE network. Stay tuned for more empowering opportunities aimed at elevating scientific excellence and leadership in research!

During the virtual mobility (VM), it was given as a 1-hour workshop about grant writing as storytelling.

The workshop was structured in two parts, both focusing on effective grant writing as a narrative practice.In the first part, it was emphasized the importance of viewing grant writing as a form of storytelling. And highlighted how this approach can help researchers communicate their projects more compellingly and persuasively to grant reviewers. The session focused on the underlying reasons why structuring research proposals in a narrative form can increase the chances of success.

The second part provided participants with a rhetorical framework designed to guide researchers in shaping the narrative of their grant proposals. It was shared practical insights, grounded in my own experience as an academic researcher, on how to craft a clear and impactful narrative that addresses the core criteria reviewers look for. This section of the workshop equipped participants with strategies for articulating their research ideas in a way that effectively communicates the value of their proposals. Moreover, supported document was shared that included some information about writing skills guidelines.

Young researchers committee hosting the section and during the workshop primarily we tried to get involved young researchers actively engaging with the content, participating in discussions, and reflecting on how these strategies could be applied to their grant writing efforts.

The workshop’s alignment with the COST Excellence and Inclusivity Policy ensured a diverse mix of participants in terms of geographical representation, career stage, and gender.

The three pages long report (p7-9) from Prof. Yesim Isil Ulman introduced the CA21139 IMPROVE together with the Istanbul Workshop of its Ethics X-over Working Group in the September 2024 Newsletter of the EACME (Home | European Association of Centres of Medical Ethics).