THREE-STAGE APPROACH FOR EVALUATION OF A CHEMICALLY DEFINED CELL CULTURE MEDIUM FOR THE CACO-2 CELL LINE: SHORT-TERM EFFECTS, DIFFERENTIATION POTENTIAL AND LONG-TERM CULTIVATION

Christian Schmidt1,2 and Joachim Wiest1,2*

11th World Congress on Alternatives and Animal Use in the Life Science, Abstract overview, 2020, 53

1 cellasys GmbH, Kronburg / Germany
2 Technical University of Munich, Heinz-Nixdorf-Chair of Biomedical Electronics, Department of Electrical and Computer Engineering, TranslaTUM, Munich / Germany

Corresponding author: wiest@cellasys.com

Finding a chemically-defined cell culture medium as a replacement for FBS is a timeconsuming and costly process. In this work, we present a three-stage approach for evaluating the chemically-defined DMEM/F12+ITS medium (SOP-G200-005_DME_F12+ITS) for the Caco-2 cell line. The three stages are increasingly lengthy and identify adhesion, proliferation and metabolism changes of the cell culture. The first stage utilizes the microphysiometric system intelligent mobile lab for in-vitro diagnostic (IMOLA-IVD) to reveal any short-term effects caused by the chemically-defined medium. The IMOLA-IVD device enables an automated cell analysis by label-free measurements of the acidification rate and impedance. Additionally, the device provides fresh nutrients to the Caco-2 cells regularly by a pre-programmable protocol. Herein, the used protocol cellasys #8 is defined as follows: 6 h DMEM + 5% FBS, 6 h DMEM/F12+ITS, 4 h DMEM + 5% FBS, 4 h DMEM/F12+ITS, 4h positive control with 2% Sodium dodecyl sulphate (SDS). In the experimental results no short-term effects and cellular stress responses are visible. This suggests that all major nutrients are present in the chemically-defined medium. The second stage is a differentiation cultivation in a T25 flask for 40 days while the third stage is a long-term cultivation for 100 days. The qualitative results of these stages obtained by a light microscope show that the proliferation and adhesion is reduced compared to DMEM + 5% FBS cultivated cells but is constant during the long-term cultivation suggesting that there are no missing nutrients. With the IMOLA-IVD system, the screening time for finding minimal, chemically-defined media can be reduced. The real-time measurement of the device shows any cell stress caused by missing nutrients in a chemically-defined medium formulation. This enables a rapid first stage screening with a duration of 24 hours.

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