3Rs IN ACTION

TENTH WOLRD CONGRESS – ALTERNATIVES AND ANIMAL USE IN THE LIFE SCIENCES, August 20-24, 2017, Seattle, Washington, USA

Joachim Wiest, Michael Feil, Tilo Weber, Roman Kolar: Chemically defined cell culture practice of L929 fibroblasts

Living cells are a promising tool to develop alternatives to animal experiments. To provide them for cell based assays e.g. cell lines are kept in CO2 incubators and split once a week with feeding approximately every third day. The guidelines for good cell culture practice give recommendations how to work in cell culture [1]. A remaining issue is that many laboratories use fetal bovine serum (FBS) in standard cell culture. Beside the questionable way of manufacturing of FBS [2], it is chemically not defined. The variations in FBS may be a driver for the reproducibility problem in biomedical research. For our L929 cell line, the cell culture media with 10% FBS was replaced by the chemically defined DME/F12 + ITS mixture [2]. With this procedure we maintained the cell line now for more than a year. Although the cells changed their morphology they are used successful in biocompatibility testing [3] and in microphysiometric experiments to determine the eye irritation potential of chemicals.

 

[1]     Coecke S, Balls M, Bowe G, Davis J, Gstraunthaler G, Hartung T, Hay R, Merten OW, Price A, Schechtman L, Stacey G, Stokes W; Second ECVAM Task Force on Good Cell Culture Practice (2005) Guidance on good cell culture practice. a report of the second ECVAM task force on good cell culture practice. Altern Lab Anim 33: 261-87

[2]     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

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

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