REVIEW article

Front. Toxicol.

Sec. In Vitro Toxicology

Volume 7 - 2025 | doi: 10.3389/ftox.2025.1612903

This article is part of the Research TopicAdvancing In Vitro Cell Culture Practices: Achieving Truly Animal-Free Experiments and Scientifically Reliable and Reproducible MethodsView all 6 articles

Fetal bovine serum: How to leave it behind in the pursuit of more reliable science

Provisionally accepted
Tilo WeberTilo Weber1*Atena Malakpour PermlidAtena Malakpour Permlid2Aline CharyAline Chary3Vito D'AlessandroVito D'Alessandro4Leah HautLeah Haut1Sebastian SeufertSebastian Seufert5Esther Veronika WenzelEsther Veronika Wenzel6James HickmanJames Hickman7Karen BiebackKaren Bieback8Joachim WiestJoachim Wiest10,9Wilhelm Gerhard DirksWilhelm Gerhard Dirks11Sandra COECKESandra COECKE12Stina OredssonStina Oredsson13
  • 1Animal Welfare Academy of the German Animal Welfare Federation, Neubiberg, Germany
  • 2Department of Health Technology, Technical University of Denmark, Lyngby, Denmark
  • 3Department of Environmental Research and Innovation, Luxembourg Institute of Science and Technology (LIST), Belval, Luxembourg, Luxembourg
  • 4Bruno Cell S.r.l., Trento, Italy
  • 5Machine Learning Expert, Zeil am Main, Germany
  • 6Abcalis GmbH, Braunschweig, Germany
  • 7NanoScience Technology Center, University of Central Florida, Orlando, United States
  • 8Institute for Transfusion Medicine and Immunology, University Medical Centre Mannheim, University of Heidelberg, Mannheim, Baden-Württemberg, Germany
  • 9cellasys know-how UG, Kronburg, Germany
  • 10Heinz-Nixdorf-Chair of Biomedical Electronics, School of Computation, Information and Technology, Technical University of Munich, TranslaTUM, Munich, Germany
  • 11Leibniz-Institute DSMZ, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
  • 12European Commission, Joint Research Centre (JRC), Ispra, Lombardia, Italy
  • 13Department of Biology, Faculty of Science, Lund University, Lund, Sweden

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Cell cultures form the backbone for scientific research and development, but also for clinical diagnostics and biotechnology. Supplying cells in vitro with growth factors, hormones, and other nutrients is achieved most often by supplementing culture media with fetal bovine serum (FBS). Despite its nearly ubiquitous use, there are major reproducibility, safety, and animal welfare issues arguing the need to replace FBS. Fortunately, numerous FBS replacements have been validated and are publicly or commercially available, making it possible to leave FBS behind. Successful examples – including serum-free, animal-component-free, and chemically defined media applications – are highlighted in this review for the cultivation of stem cells and organoids, the development of organ-on-a-chip systems, the bioprinting of tissues, and the production of cultivated meat, antibodies, and vaccines, including the conduct of cytotoxicity tests and the cryopreservation of cells. Moreover, the use of fully animal-free models and methodologies is further discussed to promote their broader acceptance and adoption within the global scientific research and development community. In this regard, this review discusses novel avenues to address the scientific and practical hurdles that might limit a full transition from FBS to fully defined cell culture media and offers a brief perspective on potential future directions.

Keywords: Animal-free in vitro, Chemically defined media, complex in vitro models, Cryopreservation, cultivated meat, Fetal bovine serum replacement, reproducibility, Xeno-free and serum-free media

Received: 16 Apr 2025; Accepted: 12 Jun 2025.

Copyright: © 2025 Weber, Malakpour Permlid, Chary, D'Alessandro, Haut, Seufert, Wenzel, Hickman, Bieback, Wiest, Dirks, COECKE and Oredsson. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Tilo Weber, Animal Welfare Academy of the German Animal Welfare Federation, Neubiberg, Germany

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Supplementary Material