University of Geneva Medical School, Institute of Genetics and Genomics in Geneva (iGE3)

2 ARTICLES PUBLISHED IN JoVE

Genetics

Parallel Measurement of Circadian Clock Gene Expression and Hormone Secretion in Human Primary Cell Cultures

Volodymyr Petrenko^*1, Camille Saini^*2, Laurent Perrin^*1, Charna Dibner¹

¹Department of Medical Specialties, Division of Endocrinology, Diabetes, Hypertension and Nutrition, Diabetes Center, University of Geneva Medical School, Institute of Genetics and Genomics in Geneva (iGE3), ²Population Epidemiology Unit (UEP), Community Medicine, Geneva University Hospital

Here, we describe settings to monitor in parallel circadian bioluminescence and the secretory activity of human islet cells and primary myotubes. For this, we employed lentiviral gene delivery of a luciferase core clock reporter, followed by in vitro synchronization and collection of outflow medium by continuous cell perifusion.

JoVE Journal

Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes

Stephanie Chanon¹, Christine Durand¹, Aurelie Vieille-Marchiset¹, Maud Robert², Charna Dibner³, Chantal Simon¹, Etienne Lefai¹

¹CarMeN Laboratory, INSERM U1060, INRA 1397, University of Lyon, ²Department of digestive and bariatric surgery, Obesity Integrated Center, University Hospital of Edouard Herriot, Hospices Civils de Lyon, Lyon 1 University, ³Division of Endocrinology, Diabetes, Hypertension and Nutrition, Department of Clinical Medicine, Faculty of Medicine, University of Geneva

In this method, human primary muscle cells are cultured in vitro to obtain differentiated myotubes and glucose uptake rates are measured. We provide a detailed protocol to quantify rates in basal and insulin-stimulated states using radiolabeled [³H] 2-deoxy-D-Glucose.