Montarras, D, Morgan, J, Collins, C, Relaix, F, Zaffran, S, Cumano, A et al.. Direct isolation of satellite cells for skeletal muscle regeneration. Science 309: 2064-2067

CNRS Unité de Recherche Associée 2578, Department of Developmental Biology, INSERM, Pasteur Institute, 75724 Paris Cedex 15, France.
Science (Impact Factor: 33.61). 10/2005; 309(5743):2064-7. DOI: 10.1126/science.1114758
Source: PubMed


Muscle satellite cells contribute to muscle regeneration. We have used a Pax3(GFP/+) mouse line to directly isolate (Pax3)(green fluorescent protein)-expressing muscle satellite cells, by flow cytometry from adult skeletal muscles, as a homogeneous population of small, nongranular, Pax7+, CD34+, CD45-, Sca1- cells. The flow cytometry parameters thus established enabled us to isolate satellite cells from wild-type muscles. Such cells, grafted into muscles of mdx nu/nu mice, contributed both to fiber repair and to the muscle satellite cell compartment. Expansion of these cells in culture before engraftment reduced their regenerative capacity.

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Available from: Terry Partridge, Oct 03, 2014
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    • "For the success of applications employing myoblasts, the ability to obtain sufficient numbers of functional myoblasts is critical, and in vitro expansion of autologous or allogeneic satellite cells is the most practical way to acquire large number of myoblasts. However , once plated on standard tissue culture plastic, the stemness, a term used to describe stem cells' self-renewal and regenerative capacity, of myoblasts is lost rapidly, yielding cells with greatly diminished regenerative potential and therapeutic potency [9] [10] [11]. "
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    ABSTRACT: The application of satellite cell-derived myoblasts in regenerative medicine has been restricted by the rapid loss of stemness during in vitro cell expansion using traditional culture systems. However, studies published in the past decade have highlighted the influence of substrate elasticity on stem cell fate and revealed that culture on a soft hydrogel substrate can promote self-renewal and prolong the regenerative potential of muscle stem cells. Whether hydrogel substrates have similar effects after long-term robust expansion remains to be determined. Herein we prepared an elastic chitosan/beta-glycerophosphate/collagen hydrogel mimicking the soft microenvironment of muscle tissues for use as the substrate for satellite cell culture and investigated its influence on long-term cell expansion. After 20 passages in culture, satellite cell-derived myoblasts cultured on our hydrogel substrate exhibited significant improvements in proliferation capability, cell viability, colony forming frequency, and potential for myogenic differentiation compared to those cultured on a routine rigid culture surface. Immunochemical staining and western blot analysis both confirmed that myoblasts cultured on the hydrogel substrate expressed higher levels of several differentiation-related markers, including Pax7, Pax3, and SSEA-1, and a lower level of MyoD compared to myoblasts cultured on rigid culture plates (all p<0.05). After transplantation into the tibialis anterior of nude mice, myoblasts that had been cultured on the hydrogel substrate demonstrated a significantly greater engraftment efficacy than those cultured on the traditional surface. Collectively, these results indicate that the elastic hydrogel substrate supported robust expansion of murine myoblasts and enhanced their engraftment in vivo. Copyright © 2015. Published by Elsevier Inc.
    Experimental Cell Research 07/2015; 337(1). DOI:10.1016/j.yexcr.2015.07.021 · 3.25 Impact Factor
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    • "Adipose stem cells for muscle regeneration reviewed in (Motohashi and Asakura, 2014). The classical stem cell from muscle is the satellite cell (Mauro, 1961; Montarras et al., 2005; Scharner and Zammit, 2011), however, in recent years several groups have described other myoprogenitors in muscle tissue such as muscle side-population cells (Gussoni et al., 1999; Asakura and Rudnicki, 2002), muscle-derived stem cells (Qu-Petersen et al., 2002), interstitial cells (Mitchell et al., 2010), and muscle-derived CD133 + stem cells (Benchaouir et al., 2007). Other cells present in muscles called fibroadipogenic progenitors have been shown to crosstalk with satellite cells to enhance myogenesis (Joe et al., 2010; Uezumi et al., 2010). "
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    ABSTRACT: Regenerative capacity of skeletal muscles resides in satellite cells, a self-renewing population of muscle cells. Several studies are investigating epigenetic mechanisms that control myogenic proliferation and differentiation to find new approaches that could boost regeneration of endogenous myogenic progenitor populations. In recent years, a lot of effort has been applied to purify, expand and manipulate adult stem cells from muscle tissue. However, this population of endogenous myogenic progenitors in adults is limited and their access is difficult and invasive. Therefore, other sources of stem cells with potential to regenerate muscles need to be examined. An excellent candidate could be a population of adult stromal cells within fat characterized by mesenchymal properties, which have been termed adipose-derived stem cells (ASCs). These progenitor adult stem cells have been successfully differentiated in vitro to osteogenic, chondrogenic, neurogenic and myogenic lineages. Autologous ASCs are multipotent and can be harvested with low morbidity; thus, they hold promise for a range of therapeutic applications. This review will summarize the use of ASCs in muscle regenerative approaches.
    Frontiers in Aging Neuroscience 07/2015; 7:123. DOI:10.3389/fnagi.2015.00123 · 4.00 Impact Factor
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    • "In such a view our data contribute to the understanding of the paracrine effects of MSC-based therapy on the host skeletal muscle and suggest that the release of this sphingolipid by the implanted cells may be a key point for recruiting host myoblasts to participate in the muscle repair. The possible clinical relevance of our findings is highlighted by the following observations: i) skeletal muscle is the largest organ of the human body and numerous traumatic injuries and pathologies can affect its functionality [44]; ii) the activity of the resident muscle stem cells, satellite cells, representing approximately 2% of the total muscle cells, is severely compromised in degenerative diseases and in volumetric muscle loss [44]–[47]; iii) satellite cells, the most obvious cell candidate to be transplanted for muscle regeneration, have some major criticisms due to their heterogeneity [48], [49], loss of stemness after culturing [50] scarce cell survival in the host tissue [51] and the inability to cross the endothelial wall [52], thus restricting their use to local application and the difficulty of obtaining large numbers of cells from a donor. Moreover, the fact that S1P is being recognized as a crucial molecule capable of regulating many fundamental skeletal muscle processes, involving not only the stimulatory action on satellite cell growth [31], [53]–[55], but also resistance to fatigue [56], regulation of muscle contraction [25], protection of the muscle fibers against injury [31], [57] may likely contribute to increase the need of MSC therapy for skeletal muscle disorders. "
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    ABSTRACT: Bone-marrow-derived mesenchymal stromal cells (MSCs) have the potential to significantly contribute to skeletal muscle healing through the secretion of paracrine factors that support proliferation and enhance participation of the endogenous muscle stem cells in the process of repair/regeneration. However, MSC-derived trophic molecules have been poorly characterized. The aim of this study was to investigate paracrine signaling effects of MSCs on skeletal myoblasts. It was found, using a biochemical and morphological approach that sphingosine 1-phosphate (S1P), a natural bioactive lipid exerting a broad range of muscle cell responses, is secreted by MSCs and represents an important factor by which these cells exert their stimulatory effects on C2C12 myoblast and satellite cell proliferation. Indeed, exposure to conditioned medium obtained from MSCs cultured in the presence of the selective sphingosine kinase inhibitor (iSK), blocked increased cell proliferation caused by the conditioned medium from untreated MSCs, and the addition of exogenous S1P in the conditioned medium from MSCs pre-treated with iSK further increased myoblast proliferation. Finally, we also demonstrated that the myoblast response to MSC-secreted vascular endothelial growth factor (VEGF) involves the release of S1P from C2C12 cells. Our data may have important implications in the optimization of cell-based strategies to promote skeletal muscle regeneration.
    PLoS ONE 09/2014; 9(9):e108662. DOI:10.1371/journal.pone.0108662 · 3.23 Impact Factor
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