Title: Skeletal muscle cells derived from human pluripotent stem cells
Legend: Suzuki lab recently initiated a new project to establish skeletal muscle progenitor/stem cells derived from human pluripotent sources. Our culture method can produce skeletal muscle progenitor cells from human pluripotent stem cells generated from both healthy donors and patients with neuromuscular disorders. Such differentiated muscle cells are available for further understanding of disease mechanisms.
(Left) Differentiating muscle cells are labeled with protein markers that can help identifying the stage of cell maturation. These muscle cells are prepared from human induced pluripotent stem cells, which were originally created from human skin cells by cell reprograming. Co-localization of three colors (red, MyoD for early myoblasts; green, Myogenin for late committed myoblasts; blue, Hoechst for cell nuclei) is visualized under a fluorescence microscope. The combination of these labels gives rise to a variety of colors, which represents a variety of the maturation stages in the plated cells and seems to simulate the timeline of human embryonic development.
(Right) Mature muscle cells are stained green (myosin heavy chain, MHC) in this micrograph of cells grown from human embryonic stem cells. Cell nuclei are stained blue with Hoechst dye. The muscle fibers contain multiple nuclei.
Citation: Hosoyama T, McGivern JV, Van Dyke JM, Ebert AD, Suzuki M. Derivation of myogenic progenitors directly from human pluripotent stem cells using a sphere-based culture. Stem Cells Translational Medicine. 3(5): 564-74, 2014.
Abstract: Using stem cells to replace degenerating muscle cells and restore lost skeletal muscle function is an attractive therapeutic strategy for treating neuromuscular diseases. Myogenic progenitors are a valuable cell type for cell-based therapy and also provide a platform for studying normal muscle development and disease mechanisms in vitro. Human pluripotent stem cells represent a valuable source of tissue for generating myogenic progenitors. Here, we present a novel protocol for deriving myogenic progenitors from human embryonic stem (hES) and induced pluripotent stem (iPS) cells using free-floating spherical culture (EZ spheres) in a defined culture medium. hES cell colonies and human iPS cell colonies were expanded in medium supplemented with high concentrations (100 ng/ml) of fibroblast growth factor-2 (FGF-2) and epidermal growth factor in which they formed EZ spheres and were passaged using a mechanical chopping method. We found myogenic progenitors in the spheres after 6 weeks of culture and multinucleated myotubes following sphere dissociation and 2 weeks of terminal differentiation. A high concentration of FGF-2 plays a critical role for myogenic differentiation and is necessary for generating myogenic progenitors from pluripotent cells cultured as EZ spheres. Importantly, EZ sphere culture produced myogenic progenitors from human iPS cells generated from both healthy donors and patients with neuromuscular disorders (including Becker’s muscular dystrophy, spinal muscular atrophy, and familial amyotrophic lateral sclerosis). Taken together, this study demonstrates a simple method for generating myogenic cells from pluripotent sources under defined conditions for potential use in disease modeling or cell-based therapies targeting skeletal muscle. PMCID: PMC4006483
About the lab: Masatoshi Suzuki’s lab is focused on developing stem cell technology for use in treating and modeling neuromuscular diseases. The team has engaged in basic and translational studies using human neural progenitor cells, mesenchymal stem cells, and pluripotent stem cells to develop therapeutic strategies for these devastating diseases.