Viability of olfactory ensheathing cells after hypoxia and serum deprivation: Implication for therapeutic transplantation

J Neurosci Res. 2014 Dec;92(12):1757-66. doi: 10.1002/jnr.23442. Epub 2014 Jun 27.

Abstract

Olfactory ensheathing cells (OECs) represent glial cells supporting neuronal turnover in the olfactory system. In vitro, OECs promote axonal growth as a source of neurotrophic growth factors; in vivo, they produce myelin, promoting remyelination of damaged axons. Consequently, OEC transplantation appears to be a promising treatment for spinal cord injury, although the functional recovery is limited. This might be ascribed to the microenvironment at the lesion site, lacking growth factors (GFs), nutrients, and oxygen. To mimic this condition, we used an in vitro approach by growing primary neonatal mouse OECs under hypoxic conditions and/or serum deprivation. In addition, we compared OECs survival/proliferation with that of primary cultures of Schwann cells (SCs) and astrocytes under the same experimental conditions. Cultures were analyzed by immunocytochemistry, and cell viability was evaluated by MTT assay. Different GFs, such as NGF, bFGF, and GDNF, and their combination were used to rescue cells from serum and/or oxygen deprivation. We show that the cell types were differently sensitive to the tested stress conditions and that OECs were the most sensitive among them. Moreover, OEC viability was rescued by bFGF under serum-deprived or hypoxic condition but not under conditions of drastic serum deprivation and hypoxia. bFGF was effective also for the other cell types, whereas the effect of the other GFs was negligible. This model suggests that administration of bFGF might be considered useful to sustain cell survival/proliferation after transplantation of OECs either alone or in combination with other glial cell types.

Keywords: glial cells; hypoxia; neuroprotection; neurotrophic factors; olfactory bulb.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cell Survival / drug effects
  • Cells, Cultured
  • Culture Media, Serum-Free / pharmacology*
  • Gene Expression Regulation / drug effects
  • Glial Fibrillary Acidic Protein
  • Hypoxia / pathology*
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Mice
  • Neuroglia / drug effects
  • Neuroglia / metabolism
  • Neuroglia / pathology*
  • Olfactory Bulb / cytology*
  • S100 Proteins / metabolism
  • Schwann Cells / drug effects
  • Schwann Cells / metabolism
  • Schwann Cells / pathology*

Substances

  • Culture Media, Serum-Free
  • Glial Fibrillary Acidic Protein
  • Hif1a protein, rat
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Intercellular Signaling Peptides and Proteins
  • S100 Proteins