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IRCCS Fondazione Santa Lucia

2 ARTICLES PUBLISHED IN JoVE

Biology

Generation of Myospheres From hESCs by Epigenetic Reprogramming

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.

Biology

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

Francesco Castagnetti^1,2, Elisabetta Fiacco³, Carol Imbriano⁴, Lucia Latella^1,2

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.