3 articles published in JoVE
Single Myofiber Isolation and Culture from a Murine Model of Emery-Dreifuss Muscular Dystrophy in Early Post-Natal Development Gloria Pegoli1,2, Federica Lucini1,2, Chiara Mozzetta3, Chiara Lanzuolo1,2,4 1Istituto Nazionale di Genetica Molecolare "Romeo ed Enrica Invernizzi", 2IRCCS Santa Lucia Foundation, 3Department of Biology and Biotechnology, "C. Darwin" Sapienza University, CNR - IBPM, 4CNR – Institute of Biomedical Technologies (ITB) Here, we propose a method to efficiently obtain single muscle fibers at early post-natal developmental stages from homozygous mutant Lamin Δ8-11 mouse model, a very severe model for Emery-Dreifuss muscular dystrophy (EDMD).
In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells Francesco Castagnetti1,2, Elisabetta Fiacco3, Carol Imbriano4, Lucia Latella1,2 1Department of Medicine, Institute of Translational Pharmacology, Italian National Research Council, 2Epigenetics and Regenerative Medicine, IRCCS Fondazione Santa Lucia, 3Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), 4Department of Life Sciences, University of Modena and Reggio Emilia Active autophagy is associated with productive muscle regeneration, which is essential for Muscle Stem Cell (MuSC) activation. Here, we provide a protocol for the in situ detection of LC3, an autophagy marker in MyoD-positive MuSCs of muscle tissue sections from control and injured mice.
Generation of Myospheres From hESCs by Epigenetic Reprogramming Sonia Albini1, Pier Lorenzo Puri1,2 1Muscle Development and Regeneration Program, Sanford-Burnham Institute for Medical Research, 2IRCCS Fondazione Santa Lucia Here, we describe a protocol based on epigenetic reprogramming of human embryonic stem cells (hESCs) toward generating a homogeneous population of skeletal muscle progenitors that under permissive culture conditions form three-dimensional clusters of contractile myofibers (myospheres), which recapitulate biological features of human skeletal muscles.