Advancing In Vitro Cell Culture Practices: Achieving Truly Animal-Free Experiments and Scientifically Reliable and Reproducible Methods

Culturing cells outside the body is integral in a vast context of areas such as basic life science research to understand molecular mechanisms involved in different remedies important for drug discovery. Additionally, it plays an important role in developmental biology, regenerative medicine, toxicological research, and assessment for evaluating compound toxicity. The ability to culture a variety of different cells outside of the body has definitely contributed to the advancement of cell biology knowledge. However, despite all these positive facts regarding cell culturing, challenges arose in 1958 when fetal bovine serum (FBS) was introduced as an omnipresent cell culture supplementation. Since then, the use of FBS has presented scientific issues related to toxicity and variability, which have gradually but too slowly come into light. These factors are probably part of the reproducibility challenges facing life science research reported lately. It is peculiar that scientists of 2024 culturing human cells use 10 % FBS in the medium without critically reflecting if the data received are reliably representative for Homo sapiens. In addition, at a time when there is a collective effort and drive towards reducing the use of animals in research from ethical points of view, the attention towards animal welfare aspects of FBS should be highlighted. Thus, there is an urgent need to get rid of and eliminate FBS in cell culturing, in fact we must get rid of all animal-derived products in cell culturing practices. The goal should be to humanize cell culturing, particularly when using human cells, to better mimic the true environment in the body.

The goal is to present and disseminate articles with easily implemented information regarding the use of non-animal products in different experimental settings involving cell lines. These articles aim to encourage and stimulate to the true use of animal-free experimental approaches. The first and main goal is to help researchers to abandon FBS but also other animal-derived sera. Additionally, we aim to supply research data that can lead to the elimination of use of Matrigel isolated from mice harboring Engelbreth-Holm-Swarm mouse tumors as well as trypsin and other animal-derived materials. All submitted research should be also without the use of live animals.
In this Research Topic, we welcome submissions of original research, reviews, mini-reviews, methods, perspectives, community case studies, conceptual analysis, data reports, policy briefs, brief research reports, general commentaries, and opinions.
Areas to be covered in this Research Topic may include but are not limited to:
• Validation of OECD test guidelines using animal-product-free media, preferably using open-source media to emphasis on advancing reproducible methodologies.
• Explore strategies for the replacement of FBS in cosmetic testing strategies. Emphasizing the importance of ethics in the cosmetic industry to making cosmetics truly animal-free.
• Replacements of Matrigel and other animal-derived hydrogels.
• Replacements of animal-derived trypsin in cell culture practices and transition to using alternative animal-free dissociation reagent sources.
• Establishment of cell lines in animal-product free media preferably using (non-proprietary) open-source media. This would address the entire cell culture process including passaging and cell banking.
• Stem cell research free of animal-derived materials.
• Reevaluation of already existing articles where the only used animal-derived product is bovine serum albumin (BSA). We would strongly urge the authors to substitute BSA with human serum albumin (HSA).