TENTH WOLRD CONGRESS – ALTERNATIVES AND ANIMAL USE IN THE LIFE SCIENCES, August 20-24, 2017, Seattle, Washington, USA
Roman Kolar, Joachim Wiest, Michael Feil, Tilo Weber: 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 . A remaining issue is that many laboratories use fetal bovine serum (FBS) in standard cell culture. Beside the questionable way of manufacturing of FBS , 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 . 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  and in microphysiometric experiments to determine the eye irritation potential of chemicals.
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