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Biology

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

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Biology

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JoVE Journal Biology

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

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

DOI:

10.3791/55743-v

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08:03 min

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June 25, 2017

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Stephanie Chanon, Christine Durand, Aurelie Vieille-Marchiset, Maud Robert, Charna Dibner, Chantal Simon, Etienne Lefai

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

Chapters

00:05Title
00:34Cell Culture Media and Solution Preparation
02:45Culture of Human Primary Muscle Cells
04:19Insulin Stimulation and Glucose Uptake
05:30Cell Lysis
06:06Determination of Radiolabeled Glucose
06:52Results: Effective Glucose Uptake Measurement and Analysis of Response to Insulin Stimulation
07:37Conclusion

Summary

Automatic Translation

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.

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