Case Studies Exemplifying the Transition to Animal Component-free Cell Culture

Abstract

Cell culture techniques are strongly connected with modern scientific laboratories and production facilities. Thus, choosing the most suitable medium for the cells involved is vital, not only directly to optimise cell viability but also indirectly to maximise the reliability of the experiments performed with the cells. Fetal bovine or calf serum (FBS or FCS, respectively) is the most commonly used cell culture medium supplement, providing various nutritional factors and macromolecules essential for cell growth. Yet, the use of FBS encompasses a number of disadvantages. Scientifically, one of the most severe disadvantages is the lot-to-lot variability of animal sera that hampers reproducibility. Therefore, transitioning from the use of these ill-defined, component-variable, inconsistent, xenogenic, ethically questionable and even potentially infectious media supplements, is key to achieving better data reproducibility and thus better science. To demonstrate that the transition to animal component-free cell culture is possible and achievable, we highlight three different scenarios and provide some case studies of each, namely: i) the adaptation of single cell lines to animal component-free culture conditions by the replacement of FBS and trypsin; ii) the adaptation of multicellular models to FBS-free conditions; and (iii) the replacement of FBS with human platelet lysate (hPL) for the generation of primary stem/stromal cell cultures for clinical purposes. By highlighting these examples, we aim to foster and support the global movement towards more consistent science and provide evidence that it is indeed possible to step out of the currently smouldering scientific reproducibility crisis.

Advances in Animal Models and Cutting-Edge Research in Alternatives: Proceedings of the Third International Conference on 3Rs Research and Progress, Vishakhapatnam, 2022

“…87 In addition, living animal bodies are used as manufacturing devices, and animals can suffer from side effects and long-lasting consequences of these procedures (another example of this is growing cancer cells in mice to produce Matrigel ® ). 90 He highlighted a number of case studies exemplifying the transition to animal component-free cell culture, 91 and listed various replacements for animal-derived reagents — for example, the use of human platelet lysate (hPL) as a supplement for human mesenchymal stem/stromal cells, and the use of xeno-free chemically defined media. 91,92 Finally, Mr Weber provided guidelines to find, 93 as well as produce, optimised serum-free cell culture media, 94 and shared details of a practical workshop on replacing FBS.…”

“…90 He highlighted a number of case studies exemplifying the transition to animal component-free cell culture, 91 and listed various replacements for animal-derived reagents — for example, the use of human platelet lysate (hPL) as a supplement for human mesenchymal stem/stromal cells, and the use of xeno-free chemically defined media. 91,92 Finally, Mr Weber provided guidelines to find, 93 as well as produce, optimised serum-free cell culture media, 94 and shared details of a practical workshop on replacing FBS. 95 He briefly touched on the replacements available for other animal-derived reagents and endotoxin/pyrogen tests, 91 hence setting the path to achieving a truly animal-free laboratory working environment, to the benefit of animals, scientists and patients.…”

A worldwide survey on the use of animal‐derived materials and reagents in scientific experimentation

“…Furthermore, an immense step forward could be achieved if peer-reviewers would demand to verify experimental results in a xeno-free or chemically defined environment [47]. Such a step is in the very own interest of reviewers, applicants and journals as it can ensure reproducibility for published results.…”

Editorial

“…Serum-free approaches are further explored in the article by Weber et al, 2 who demonstrate, by providing a range of examples, that the transition to animal component-free cell culture is possible and achievable. They highlight three different scenarios and provide case studies for each.…”