The SH-SY5Y neuronal control cells and mutant cybrid cells carrying m.3243A > G with 98%, 90%, and 70% mutation loads and 143B osteosarcoma cybrid control and mutant cells were cultured in standard DMEM high glucose media (4.5 g/L) or in low glucose (0.5 g/L) (PAN biotech, Aidenbach, Germany), supplemented with 10% foetal bovine serum (PAN biotech), 1% glutamine, and 50 μg/mL uridine (Sigma-Aldrich, Saint-Louis, MO, USA) at 37 °C in the presence of 5% CO2 as described elsewhere [16 (link)]. Control and mutant cells were exposed to 5 mM of acetoacetate and β-D-hydroxybutyrate (Sigma-Aldrich, Saint-Louis, MI, USA) in a low glucose medium for 48 h. Control cells were exposed to rotenone (200 nM) a complex I inhibitor for 15 h.
Belal S., Goudenège D., Bocca C., Dumont F., Chao De La Barca J.M., Desquiret-Dumas V., Gueguen N., Geffroy G., Benyahia R., Kane S., Khiati S., Bris C., Aranyi T., Stockholm D., Inisan A., Renaud A., Barth M., Simard G., Reynier P., Letournel F., Lenaers G., Bonneau D., Chevrollier A, & Procaccio V. (2022). Glutamate-Induced Deregulation of Krebs Cycle in Mitochondrial Encephalopathy Lactic Acidosis Syndrome Stroke-Like Episodes (MELAS) Syndrome Is Alleviated by Ketone Body Exposure. Biomedicines, 10(7), 1665.
Corresponding Organization : Centre National de la Recherche Scientifique
Other organizations :
Université Paris-Saclay, Laboratory Signaling and Cardiovascular Pathophysiology, Institute of Molecular Life Sciences, Semmelweis University, Université Paris Sciences et Lettres, École Pratique des Hautes Études, Sorbonne Université, Centre de Recherche Saint-Antoine, Centre Hospitalier Universitaire d'Angers
Glucose concentration in the medium (4.5 g/L or 0.5 g/L)
Exposure to acetoacetate and β-D-hydroxybutyrate (5 mM)
Exposure to rotenone (200 nM)
dependent variables
Cell growth and metabolism of SH-SY5Y neuronal control cells and mutant cybrid cells carrying m.3243A > G with 98%, 90%, and 70% mutation loads
Cell growth and metabolism of 143B osteosarcoma cybrid control and mutant cells
control variables
Cell culture conditions (37 °C, 5% CO2)
Culture medium (DMEM supplemented with 10% foetal bovine serum, 1% glutamine, and 50 μg/mL uridine)
controls
Positive control: 143B osteosarcoma cybrid control cells
Negative control: Rotenone-treated SH-SY5Y neuronal control cells
Annotations
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