The Influence of Popular Antibiotics in Culture Medium on the Cell Motility and Superoxide Production of Mesenchymal Stem Cells

Antibiotics have become an integral part of routine cell culturing. Penicillin-streptomycin mixture and gentamicin are widely used to prevent bacterial contamination of cells. A lot of publications have elucidated the effect of these antibiotics in culture medium on differentiation and metabolism of mesenchymal stem cells (MSCs). However, much less is known about their influence on cell migration and mitochondrial superoxide production. In this study, MSCs from human adipose tissue were cultured in media containing mostly used concentrations of mentioned antibiotics. In total, two concentrations of penicillin-streptomycin (100 U·ml−1–100 µg·ml−1, 200 U·ml−1–200 µg·ml−1) and gentamicin (10 µg·ml−1, 50 µg·ml−1) were tested. The acquisition of microscopic images was performed using Leica TCS SP8 X confocal laser scanning microscope. The MitoSOX Red indicator was used to observe mitochondrial superoxide. Image processing and statistical analysis was performed using MATLAB software. We found that the addition of these antibiotics to the medium negatively affects the cell migration velocity in a dose-dependent manner, with penicillin-streptomycin inhibiting cell migration less than gentamicin. Also, penicillin-streptomycin and gentamicin induce mitochondrial superoxide production in cells.