Nestin-GFP transgene reveals neural precursor cells in adult skeletal muscle

• Published in: PloS one 2011-02, Vol.6 (2), p.e16816-e16816
• Main Authors: Birbrair, Alexander, Wang, Zhong-Min, Messi, Maria Laura, Enikolopov, Grigori N, Delbono, Osvaldo
• Format: Article
• Language: English
• Subjects: Actin; Aging - metabolism; Aging - physiology; Animal tissues; Animals; Biology; Biopsy; Brain research; Cell culture; Cell Differentiation - genetics; Cell Differentiation - physiology; Cell survival; Cell Tracking - methods; Cells (biology); Cells, Cultured; Central nervous system; Degenerative diseases; Ethics; Female; Flow Cytometry; Fluorescence; Genetic engineering; Gerontology; Glutamate; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Green Fluorescent Proteins - physiology; Immunology; Intermediate filament proteins; Intermediate Filament Proteins - genetics; Intermediate Filament Proteins - metabolism; Intermediate Filament Proteins - physiology; Internal medicine; Lesions; Male; Markers; Medical research; Medicine; Mesenchyme; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muscle proteins; Muscle, Skeletal - cytology; Muscle, Skeletal - metabolism; Musculoskeletal system; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Nerve Tissue Proteins - physiology; Nervous system; Nestin; Neural stem cells; Neural Stem Cells - cytology; Neural Stem Cells - metabolism; Neural Stem Cells - physiology; Neurogenesis; Neurosciences; Precursors; Proteins; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Recombinant Fusion Proteins - physiology; Regulatory sequences; Rodents; Skeletal muscle; Smooth muscle; Stem cells; Therapy; Transgenes; Transgenes - physiology; Transgenic mice; Transplants & implants; Vascular endothelial growth factor
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0016816

Therapy for neural lesions or degenerative diseases relies mainly on finding transplantable active precursor cells. Identifying them in peripheral tissues accessible for biopsy, outside the central nervous system, would circumvent the serious immunological and ethical concerns impeding cell therapy. In this study, we isolated neural progenitor cells in cultured adult skeletal muscle from transgenic mice in which nestin regulatory elements control GFP expression. These cells also expressed the early neural marker Tuj1 and light and heavy neurofilament but not S100β, indicating that they express typical neural but not Schwann cell markers. GFP+/Tuj1+ cells were also negative for the endothelial and pericyte markers CD31 and α-smooth muscle actin, respectively. We established their a) functional response to glutamate in patch-clamp recordings; b) interstitial mesenchymal origin; c) replicative capacity; and d) the environment necessary for their survival after fluorescence-activated cell sorting. We propose that the decline in nestin-GFP expression in muscle progenitor cells and its persistence in neural precursor cells in muscle cultures provide an invaluable tool for isolating a population of predifferentiated neural cells with therapeutic potential.