Simple derivation of skeletal muscle from human pluripotent stem cells using temperature‐sensitive Sendai virus vector

• Published in: Journal of cellular and molecular medicine 2021-10, Vol.25 (20), p.9586-9596
• Main Authors: Tan, Ghee Wan, Kondo, Takayuki, Imamura, Keiko, Suga, Mika, Enami, Takako, Nagahashi, Ayako, Tsukita, Kayoko, Inoue, Ikuyo, Kawaguchi, Jitsutaro, Shu, Tsugumine, Inoue, Haruhisa
• Format: Article
• Language: English
• Subjects: Antibodies; Biotechnology; Calcium - metabolism; Calcium Signaling; Cell differentiation; Cell Differentiation - genetics; Cells, Cultured; Cellular Reprogramming - genetics; Cytology; Cytotoxicity; differentiation method; disease modelling; Electrical stimuli; Embryo cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Fluorescent Antibody Technique; Gene Expression; Genes; Genetic Vectors - genetics; High temperature; high temperature treatment; human embryonic stem cells; Humans; human‐induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Infections; Muscle Development - genetics; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - metabolism; Musculoskeletal system; Myoblasts; Myod1; Myosin; Neuromuscular diseases; Original; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Regenerative medicine; RNA polymerase; Sendai virus; Sendai virus - genetics; Skeletal muscle; Stem cells; Temperature; Transgenes
• ISSN: 1582-1838; 1582-4934
• DOI: 10.1111/jcmm.16899

Human pluripotent stem cells have the potential to differentiate into various cell types including skeletal muscles (SkM), and they are applied to regenerative medicine or in vitro modelling for intractable diseases. A simple differentiation method is required for SkM cells to accelerate neuromuscular disease studies. Here, we established a simple method to convert human pluripotent stem cells into SkM cells by using temperature‐sensitive Sendai virus (SeV) vector encoding myoblast determination protein 1 (SeV‐Myod1), a myogenic master transcription factor. SeV‐Myod1 treatment converted human embryonic stem cells (ESCs) into SkM cells, which expressed SkM markers including myosin heavy chain (MHC). We then removed the SeV vector by temporal treatment at a high temperature of 38℃, which also accelerated mesodermal differentiation, and found that SkM cells exhibited fibre‐like morphology. Finally, after removal of the residual human ESCs by pluripotent stem cell‐targeting delivery of cytotoxic compound, we generated SkM cells with 80% MHC positivity and responsiveness to electrical stimulation. This simple method for myogenic differentiation was applicable to human‐induced pluripotent stem cells and will be beneficial for investigations of disease mechanisms and drug discovery in the future.