Abstract
Muscle stem cells (MuSCs) exhibit robust myogenic potential in vivo, thus providing a promising curative treatment for muscle disorders. Ex vivo expansion of adult MuSCs is highly desired to achieve a therapeutic cell dose because of their scarcity in limited muscle biopsies. Sorting of pure MuSCs is generally required for all the current culture systems. Here we developed a soft three‐dimensional (3D) salmon fibrin gel culture system that can selectively expand mouse MuSCs from bulk skeletal muscle preparations without cell sorting and faithfully maintain their regenerative capacity in culture. Our study established a novel platform for convenient ex vivo expansion of MuSCs, thus greatly advancing stem cell‐based therapies for various muscle disorders. Stem Cells Translational Medicine 2017;6:1412–1423
Highlights
Skeletal muscle is a form of striated muscle tissue involved in diverse bodily functions
Due to the 3D microarchitecture niche of Muscle stem cells (MuSCs) in the body, we hypothesized that a 3D culture system would facilitate ex vivo expansion of MuSCs
When using fibrin gel to culture the early passage primary skeletal muscle cells that consist mainly of myoblasts and fibroblasts, we found that a low concentration (0.5 mg/ ml) of 3D salmon fibrin gel favored growth of round myoblasts even in fibroblast-favorable Dulbecco’s Modified Eagle Medium (DMEM)/F10 containing growth medium (Supporting Information Fig. S1)
Summary
Skeletal muscle is a form of striated muscle tissue involved in diverse bodily functions. The functional unit of skeletal muscle is the multinucleated myofibers formed by the fusion of hundreds of myoblasts in a process called myogenesis. Because satellite cells can self-renew [2] and differentiate into functional progeny [3, 4], there is a consensus that they are bona fide adult muscle stem cells (MuSCs). Quiescent MuSCs are characterized by their expression of Pax and/or Myf but not MyoD and myogenin (MyoG) [7]. Upon muscle injury or detachment from basal lamina, MuSCs become activated and initiate rapid expression of myogenic transcription factor MyoD, which enables cells to re-enter the cell cycle and proliferate [8,9,10]. Proliferating satellite cells and their progeny are often referred to as myogenic precursor cells. An intermediate ratio of Pax to MyoD keeps satellite cells proliferating without differentiation, while a low Pax7-to-MyoD ratio drives stem cells into terminal differentiation [7]
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