Abstract
BackgroundAdipose tissue is an excellent source for isolation of stem cells for treating various clinical conditions including injuries to the neuromuscular system. Many previous studies have focused on differentiating these adipose stem cells (ASCs) towards a Schwann cell-like phenotype (dASCs), which can enhance axon regeneration and reduce muscle atrophy. However, the stromal vascular fraction (SVF), from which the ASCs are derived, also exerts broad regenerative potential and might provide a faster route to clinical translation of the cell therapies for treatment of neuromuscular disorders.MethodsThe aim of this study was to establish the effects of SVF cells on the proliferation and differentiation of myoblasts using indirect co-culture experiments. A Growth Factor PCR Array was used to compare the secretomes of SVF and dASCs, and the downstream signaling pathways were investigated.ResultsSVF cells, unlike culture-expanded dASCs, expressed and secreted hepatocyte growth factor (HGF) at concentrations sufficient to enhance the proliferation of myoblasts. Pharmacological inhibitor studies revealed that the signal is mediated via ERK1/2 phosphorylation and that the effect is significantly reduced by the addition of 100 pM Norleual, a specific HGF inhibitor. When myoblasts were differentiated into multinucleated myotubes, the SVF cells reduced the expression levels of fast-type myosin heavy chain (MyHC2) suggesting an inhibition of the differentiation process.ConclusionsIn summary, this study shows the importance of HGF as a mediator of the SVF effects on myoblasts and provides further evidence for the importance of the secretome in cell therapy and regenerative medicine applications.
Highlights
Adipose tissue is an excellent source for isolation of stem cells for treating various clinical conditions including injuries to the neuromuscular system
stromal vascular fraction (SVF) cells enhance the proliferation of myoblasts but not via an acetylcholine-dependent mechanism The enhanced proliferation of myoblasts in the coculture experiments was blocked in the presence of a MAP kinase kinase (MEK) inhibitor, which suggested that the effect was mediated via the MAPK/Extracellular signal-regulated kinase (ERK) pathway (Fig. 2a)
Compared with the Differentiated adipose stem cell (dASC), the SVF cells expressed significantly lower levels of choline acetyltransferase (ChAT), an enzyme which is necessary for ACh production (Fig. 2b)
Summary
Adipose tissue is an excellent source for isolation of stem cells for treating various clinical conditions including injuries to the neuromuscular system. Many previous studies have focused on differentiating these adipose stem cells (ASCs) towards a Schwann cell-like phenotype (dASCs), which can enhance axon regeneration and reduce muscle atrophy. There is an activation of intrinsic satellite cells, which is a sign of neomyogenesis Despite this endogenous response, the absence of innervation leads to a Mesenchymal stem cells (MSCs) have great clinical potential to treat many human diseases and regenerate damaged tissue due to their anti-inflammatory, antiscarring, angiogenic, and pro-healing functions [3, 4]. Adipose tissue is an excellent source for isolation of MSCs [5], and previous studies have shown how adipose derived stem cells (ASCs) have pro-regenerative effects in the injured peripheral nervous system [6,7,8]. We reported that the dASCs enhanced the proliferation of myoblasts in vitro through paracrine
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