Generation of High-Yield, Functional Oligodendrocytes from a c- myc Immortalized Neural Cell Line, Endowed with Staminal Properties

Int J Mol Sci. 2021 Jan 23;22(3):1124. doi: 10.3390/ijms22031124.

Abstract

Neural stem cells represent a powerful tool to study molecules involved in pathophysiology of Nervous System and to discover new drugs. Although they can be cultured and expanded in vitro as a primary culture, their use is hampered by their heterogeneity and by the cost and time needed for their preparation. Here we report that mes-c-myc A1 cells (A1), a neural cell line, is endowed with staminal properties. Undifferentiated/proliferating and differentiated/non-proliferating A1 cells are able to generate neurospheres (Ns) in which gene expression parallels the original differentiation status. In fact, Ns derived from undifferentiated A1 cells express higher levels of Nestin, Kruppel-like factor 4 (Klf4) and glial fibrillary protein (GFAP), markers of stemness, while those obtained from differentiated A1 cells show higher levels of the neuronal marker beta III tubulin. Interestingly, Ns differentiation, by Epidermal Growth Factors (EGF) and Fibroblast Growth Factor 2 (bFGF) withdrawal, generates oligodendrocytes at high-yield as shown by the expression of markers, Galactosylceramidase (Gal-C) Neuron-Glial antigen 2 (NG2), Receptor-Interacting Protein (RIP) and Myelin Basic Protein (MBP). Finally, upon co-culture, Ns-A1-derived oligodendrocytes cause a redistribution of contactin-associated protein (Caspr/paranodin) protein on neuronal cells, as primary oligodendrocytes cultures, suggesting that they are able to form compact myelin. Thus, Ns-A1-derived oligodendrocytes may represent a time-saving and low-cost tool to study the pathophysiology of oligodendrocytes and to test new drugs.

Keywords: cell culture; immortalization; myelin; neural stem cell; neuron; oligodendrocyte.

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Adhesion Molecules, Neuronal / metabolism
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology
  • Cell Line
  • Coculture Techniques
  • Epidermal Growth Factor / pharmacology
  • Fibroblast Growth Factor 2 / pharmacology
  • Fluorescent Antibody Technique / methods
  • Gene Expression Regulation
  • Genes, myc
  • Glial Fibrillary Acidic Protein / genetics
  • Kruppel-Like Factor 4
  • Mice
  • Myelin Basic Protein / metabolism
  • Myelin Sheath / metabolism
  • Neural Stem Cells / cytology
  • Oligodendroglia / cytology*
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Biomarkers
  • Cell Adhesion Molecules, Neuronal
  • Cntnap1 protein, mouse
  • Glial Fibrillary Acidic Protein
  • Klf4 protein, mouse
  • Klf4 protein, rat
  • Kruppel-Like Factor 4
  • Mbp protein, mouse
  • Myelin Basic Protein
  • glial fibrillary astrocytic protein, mouse
  • Fibroblast Growth Factor 2
  • Epidermal Growth Factor