Chemical media as alternative to FBS
"There are several reasons to reconsider its use", said Dr. Peter Frost, CEO PELOBiotech: The harvesting process for FBS raises ethical concerns that remain unchanged since its inception. "It is crucial to explore and adopt more ethical alternatives to FBS in research", said Dr. Lothar Steeb, CEO/CSO PELOBiotech.
"A potential solution to these challenges is the use of defined media”, said Dr. Lothar Steeb, CEO/CSO PELOBiotech. Not all defined media are free of animal components. “To fully address the ethical and scientific concerns associated with FBS and animal-derived components, we need to implement defined media that are both serum-free and animal component-free”, said Dr. Peter Frost, CEO PELOBiotech. Although these products are currently available, they are often more expensive than regular media containing animal components.
FBS was first used in cell culture in 1958 by Theodore Puck, an American geneticist. "Despite numerous advances in science and technology, it remains in use", pointed out Prof. Dr. Gerhard Gstraunthaler emer, University of Innsbruck, Austria. How have FBS and other old cell culture techniques, like using stiff surfaces for cell growth, persisted through decades of rapid innovation and evolution in the field? In an era where sustainability is crucial and reducing animal consumption is a priority, "it seems paradoxical that we still rely on puncturing the hearts of unborn fetuses to grow our cells", said Prof. Dr. Gerhard Gstraunthaler, University of Innsbruck, Austria, in his lecture.
Cruelty-free animal-free research
There are many unknown factors in the FBS production process: A significant volume of fetal blood is pooled from numerous cows at various stages of pregnancy to meet global requests*(1). According to estimates, the global demand for FBS is approximately 700,000 liters annually, with production slightly exceeding this at around 800,000 liters to ensure supply. Overall, there are considerable geographical differences in the production process and quality of FBS, leading to variability in pricing, according to the International Serum Industry Association. The serum itself presents a black box: serum contains growth factors, hormones, proteins, coagulation factors, and other components, but the suppliers don’t know what the growth of the cells is in all of these is the key or triggering factor. And so the experiments might become a black box too, some work, some not, reproducibility often at stake.
“Extensive research is needed to understand the key factors in serum that trigger cell growth”, concluded Prof. Dr. rer. nat. Bernd Giebel, University Hospital Essen. By decoding these components, science can move toward developing artificial serum or other FBS alternatives that provide comparable results and are more cost-effective than currently defined media options. Prof. Dr. rer. nat. Bernd Giebel, University Hospital Essen, highlighted the challenge of high protein content in hPL, hindering the purification of extracellular vehicles (EVs). The high protein content interferes with the purification processes and contaminates EV preparations, complicating downstream analyses. Additionally, despite being derived from human waste products, human serum remains a complex mixture, or black box, with similar uncertainties to FBS, pointed out Giebel in his lecture.
More and more scientists become aware of this transition to a more sustainable, reproducible, and animal-free alternative. But how, what could be a first step? An answer, scientists could find in Gstraunthaler´'s lecture: A start could be small, just by optimizing our protocols to use a lower percentage of FBS reducing from the typical 10% to 4 or 5%. This gradual reduction allows observing how FBS concentration changes affect cell proliferation and functionality, suggested Gstraunthaler. At the conference applications were shown, so scientists could take home practical out-of-the-lab tips: “Transitioning from FBS to alternatives such as human serum or defined media is challenging and may not work for all protocols or cell types”, explained Dr. rer. nat. Anita Wagner, Technical University of Munich, how important gradual changes and optimization are to ensure successful implementation and adaptation of new methods.
"FBS is a cost-effective protein source, the rising preference for plant-based diets raises concerns about its future availability and ethical implications. As plant-based diets become more popular for environmental and ethical reasons, the demand for alternatives to FBS is expected to increase, impacting its availability*(3). "When cows are slaughtered solely for FBS production this might lead to significantly higher prices", said Dr. Peter Frost, CEO PELOBiotech. Alternatives that currently seem expensive might then become real alternatives.
Ethical and sustainable cell culture practices
"We aim to make these alternatives cost-effective which is crucial for the future of cell culture research", promised Dr. Lothar Steeb, CEO/CSO PELOBiotech. He has worked in this field for over 25 years, refining media and finding the best solution for alternatives. Achieving this will require increased demand and higher production volumes. The Cell Culture expert has already seen much greater awareness and understanding within the scientific community about the ethical issues associated with the use of FBS and other animal-derived components, "We need and are willing to take proactive steps towards developing alternatives”, assured Dr. Peter Frost, CEO PELOBiotech.
The shift from 2D to 3D cell culture marked a significant transformation in this field, and "the steady evolution continues today, it is needed for real alternatives to FBS," concluded Dr. Peter Frost, CEO PELOBiotech.
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The panel of experts
Prof. Dr. rer. nat. Bernd Giebel, University Hospital Essen, Germany, provided insights into extracellular vesicles (EVs) and the immunomodulatory effects of MSC-EVs.
Prof. Dr. Gerhard Gstraunthaler, emer., University of Innsbruck, Austria, talked about the alternatives to FBS
Prof. Dr. Kurt Pfannkuche, University of Cologne, guided attendees on cardiac organoid characterization and scalable generation.
Dr. Flávia Rezende, Center for Molecular Medicine, Goethe University in Frankfurt am Main, offered insights into using metabolomics to understand endothelial cell metabolomic cues better.
Dr. Raquel Sousa, Research and Development Scientist, abc biopply, Switzerland, showed the revolutionizing human disease models with 3D CoSeedis™ humanized multi-organoid in chip technology.
Dr. Lothar Steeb, CSO/CEO PELOBiotech GmbH, explained the alternatives of serum-free to defined media
Dr. rer. nat. Anita Wagner, Technical University of Munich, raised awareness about the risks of mitochondrial dysfunction associated with increased BMI and unhealthy lifestyles.
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References:
- *Wei, Z., Batagov, A. O., Carter, D. R. F., & Krichevsky, A. M. (2016). Fetal Bovine Serum RNA interferes with cell-culture-derived extracellular RNA. Scientific Reports. DOI: 10.1038/srep31175.
- International Serum Industry Association. (2024). FBS Production and Testing. Retrieved from org.
- Hadi, J.; Brightwell, G. Safety of Alternative Proteins: Technological, Environmental and Regulatory Aspects of Cultured Meat, Plant-Based Meat, Insect Protein and Single-Cell Protein. Foods 2021, 10, 1226. https://doi.org/10.3390/foods10061226
- Bieback, K., Hecker, A., Kocaömer, A., et al. (2009). Human alternatives to fetal bovine serum for the expansion of mesenchymal stromal cells from bone marrow. Stem Cells, 27, 2331–2341. DOI: 10.1002/stem.139
- Doucet, C., Ernou, I., Zhang, Y., et al. (2005). Platelet lysates promote mesenchymal stem cell expansion: a safety substitute for animal serum in cell-based therapy applications. Journal of Cellular Physiology, 205, 228–236. DOI: 10.1002/jcp.20327
- Gstraunthaler, G. (2003). Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX, 20, 275–281. PubMed
- van der Valk, J., Brunner, D., De Smet, K., et al. (2010). Optimization of chemically defined cell culture media. Replacing fetal bovine serum in mammalian in vitro methods. Toxicology in Vitro, 24, 1053–1063. DOI: 10.1016/j.tiv.2010.03.016
