Induction of Neuronal Differentiation of Rat Muscle-Derived Stem Cells in Vitro Using Basic Fibroblast Growth Factor and Ethosuximide

Abstract

Several studies have demonstrated that basic fibroblast growth factor (bFGF) can induce neural differentiation of mesenchymal stem cells. In this study, we investigated the neural differentiation of muscle-derived stem cells (MDSCs) following treatment with bFGF and ethosuximide, a small molecule used as an anticonvulsant in humans. Stem cells isolated from rat skeletal muscle (rMDSCs) were pre-induced by culturing with 25 ng/mL bFGF for 24 h and then were transferred to a medium supplemented with or without 4 mM ethosuximide. Neuronal differentiation was assessed by immunocytochemical and western blotting analyses of marker expression. Immunocytochemistry of rMDSCs treated with bFGF and ethosuximide identified abundant cells expressing neuronal markers (TuJ1, neuron-specific class III β-tubulin; NeuN, neuronal nuclear antigen; and NF-MH; neurofilament M and H). Olig2 (oligodendrocyte transcription factor 2)-positive cells were also observed, indicating the presence of oligodendrocyte lineage cells. These findings were substantiated by western blotting analysis of marker proteins. In particular, the expression of NeuN and TuJ1 was significantly higher in rMDSCs treated with ethosuximide and bFGF than in cells stimulated with bFGF alone (NeuN, p < 0.05 and TuJ1, p < 0.001). Expression of the astrocyte marker GFAP (glial fibrillary acidic protein) was not detected in this study. Collectively, the results showed that treatment with bFGF and ethosuximide induced effective transdifferentiation of rMDSCs into cells with a neural-like phenotype. Notably, rMDSCs treated with a combination of bFGF plus ethosuximide showed enhanced differentiation compared with cells treated with bFGF alone, implying that ethosuximide may stimulate neuronal differentiation.

Aberrant Oligodendrogenesis in Down Syndrome: Shift in Gliogenesis?

“…Donepezil was shown to promote the differentiation of primary NSCs into mature oligodendrocytes at the expense of astrocytes [150], and it was also found to enhance myelin sheath generation in neuron/glia co-cultures [151]. The FDA-approved anti-seizure drug ethosuximide was described to be capable of inducing trans-differentiation of muscle-derived stem cells into Olig2-positive oligodendroglial cells [152]. Whether ethosuximide activity could also be used to re-establish the glial cell balance needs to be shown, keeping in mind that Olig2 expression itself is not restricted to oligodendroglial cells, but is also found in the cytoplasm of astroglia [153].…”

Plasticity of Mesenchymal Stem Cells from Mouse Bone Marrow in the Presence of Conditioned Medium of the Facial Nerve and Fibroblast Growth Factor-2

“…These changes were found in most cells of group 4. Kang et al [ 22 ] investigated the neural differentiation of stem cells derived from muscle followed by treatment with an anticonvulsant and FGF. The morphological changes observed in cultures stimulated with FGF for 24 hours were consistent with GABA neurons.…”

Intrinsic Ability of Adult Stem Cell in Skeletal Muscle: An Effective and Replenishable Resource to the Establishment of Pluripotent Stem Cells

“…Kwon et al reported that valproic acid (VA), a histone deacetylase inhibitor used in the treatment of epilepsy, bipolar disorder, led to differentiation of muscle-derived stem cells toward neuronal lineages. Recently, Kang et al demonstrated that fibroblast growth factor (FGF) and ethosuximide, which is used clinically to treat absence seizures in humans, induced neuronal differentiation of muscle-derived stem cells by showing immunohistochemically positive cells for TuJ1, NeuN, and neurofilaments M and H [150]. Further studies to reveal the mechanistic background of the conversion into the neuronal lineage may reveal further potential applications of muscle-derived stem cells, which can easily regenerate and consequently be rapidly expanded ex vivo , for repair of neuronal injuries and treatment of neuronal disorders.…”