Chemically defined cell culture media – a contribution to address the reproducibility crisis in biomedical sciences

Tilo Weber1, Kristina Wagner1, Joachim Wiest2*

ALTEX Proceedings, 2021, 9, 1, 262

1 German Animal Welfare Federation
2 cellasys GmbH

Corresponding author:

The use of fetal bovine serum (FBS) in cell culture media has different drawbacks. The
harvesting of FBS from bovine fetuses after slaughter of the pregnant parent (dam)
raises ethical and legal concerns. From a pure scientific point of view the use of FBS is
inacceptable since regional differences in type and concentration of ingredients exist
(Baker, 2016, van der Valk et al., 2010). Hence, a non-definable quality of FBS
undermines data reliability and decrease or even prevent experimental reproducibility.
However, the use of chemically defined cell culture media is still scarce.
The introduction of chemically defined cell culture media is a contribution to fight the
reproducibility crisis in the biomedical sciences and an approach to address animal
welfare concerns. Usage of chemically defined medium will eliminate some unknowns in
cell culture experiments. A procedure to develop serum free media was introduced by
van der Valk and colleagues (van der Valk et al., 2010). In the presented work, a detailed
recipe to prepare a defined DMEM / Ham’s F12 + ITS cell culture medium is given. This
medium has proven to work in our laboratory for L929 and Caco-2 cell lines in
combination with a certain plastic ware (Greiner Bio-One, Advanced-TC). To prepare the
chemically defined cell culture medium (DME /F12+ITS) mix 50% DMEM (e.g. Sigma
Aldrich, D5648) and 50% Ham’s F12 (e.g. Sigma Aldrich, N6760), add 14.7 mmol/l NaCl,
20.9 mmol/l NaCHO3 and 5 ml/l ITS (Sigma Aldrich, I3146) (cellasys, 2019). The medium
was developed for cell culture in a 5% CO2 incubator. The presented method was
employed to develop a chemically defined cell-based assay for cytotoxicity
determination according to ISO10993-5 (Wiest, 2017), furnishing a first proof of its
applicability as an alternative to cell culture media containing FBS.

Baker, M. (2016). Reproducibility: Respect your cells!. Nature 537, 433–435.

cellasys (2019).

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(4), 1053–1063. doi:10.1016/j.tiv.2010.03.016;

van der Valk, J., Bieback, K., Buta, C. et al. (2018). Fetal Bovine Serum (FBS): Past – Present –
Future. ALTEX 35(1), 99–118. doi:10.14573/altex.1705101;

Wiest, J. (2017). Chemisch definiert – ein zellbasierter Zytotoxizitätsassay ohne fötales Kälberserum. BIOspektrum
23, 61. doi:10.1007/s12268-017-0768-6

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