Accedi

Cure Congenital Muscular Dystrophy

2 ARTICLES PUBLISHED IN JoVE

Biology

Tissue Triage and Freezing for Models of Skeletal Muscle Disease

Hui Meng¹, Paul M.L. Janssen², Robert W. Grange³, Lin Yang⁴, Alan H. Beggs⁵, Lindsay C. Swanson⁵, Stacy A. Cossette^1,6, Alison Frase⁷, Martin K. Childers⁸, Henk Granzier⁹, Emanuela Gussoni⁵, Michael W. Lawlor¹

¹Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, ²Department of Physiology and Cell Biology, The Ohio State University, ³Department of Human Nutrition, Foods and Exercise, Virginia Tech, ⁴Division of Biomedical Informatics, Department of Biostatistics, Department of Computer Science, University of Kentucky, ⁵Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School, ⁶Cure Congenital Muscular Dystrophy, ⁷Joshua Frase Foundation, ⁸Department of Rehabilitation Medicine, University of Washington, ⁹Department of Physiology, University of Arizona

The analysis of skeletal muscle tissues to determine structural, functional, and biochemical properties is greatly facilitated by appropriate preparation. This protocol describes appropriate methods to prepare skeletal muscle tissue for a broad range of phenotyping studies.

Medicine

Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling

Jerome D. Robin¹, Woody E. Wright¹, Yaqun Zou², Stacy A. Cossette³, Michael W. Lawlor³, Emanuela Gussoni⁴

¹Department of Cell Biology, UT Southwestern Medical Center, ²National Institute of Neurological Disorders and Stroke, National Institute of Health, ³Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, ⁴Division of Genetics and Genomics, Boston Children's Hospital

This protocol describes techniques for live cell isolation and primary culture of myogenic and fibroblast cell lines from muscle or skin tissue. A technique for the immortalization of these cell lines is also described. Altogether, these protocols provide a reliable tool to generate and preserve patient-derived cells for downstream applications.