Neurosphere cell differentiation to aldynoglia promoted by olfactory ensheathing cell conditioned medium

Abstract

Cells from central nervous system with morphology similar to radial glia and properties intermediate between astrocytes and Schwann cells were called growth-promoting glia or aldynoglia. These cells are present in adult brain olfactory bulb, hypothalamus, hypophysis, pineal gland and in the developing brain, and spinal cord (Cameron and Rakic (1991) Glia 4:124-137; Gudiño-Cabrera and Nieto-Sampedro (2000) 30:49-63). We report now that neurosphere cells, abundantly generated from E15 rat or E13 mouse corpus striatum, differentiate to aldynoglia-like cells when plated onto an adhesive substrate, and cultured in the presence of olfactory ensheathing cell conditioned medium, supplemented with EGF and bFGF. The immunophenotype, mRNA expression (microarray and RT-PCR analysis), neurite growth-promoting and ensheathing properties of the cells derived from neurospheres were similar to those of aldynoglia. Neurosphere-derived, aldynoglia-like cells expressed, with respect to neurospheres, very increased levels of GFAP, high levels of nestin and vimentin, extracellular matrix proteins, and inhibitors of the catabolism of those extracellular matrix proteins.

Increased p75 neurotrophin receptor expression in the canine distemper virus model of multiple sclerosis identifies aldynoglial schwann cells that emerge in response to axonal damage

“…This discrepancy between in vitro and in vivo findings could be due to several reasons. Since CNS Schwannlike cells have been so far only partially characterized in vitro (Donzel-P erez et al, 2009;Orlando et al, 2008), it is not clear whether the used purification protocol supported the isolation of a representative cell population. However, the fact that not a single CDV-positive Schwann-like cell expressing p75 NTR was found in vivo clearly argues against this notion.…”

Neural differentiation of transplanted neural stem cells in a rat model of striatal lacunar infarction: light and electron microscopic observations

“…We have shown in culture that neurosphere-derived cells are able to differentiate into a type of axonal-growth-promoting ensheathing glia, named aldynoglia (Gudino-Cabrera and Nieto-Sampedro, 2000; Doncel-Pérez et al, 2009). As such glia and neurons positive for GFP expression (Figure 3) facilitated the tracking of transplanted cells into the brain.…”

“…Striatum from C57BL/6 mice, transgenic for the GFP gene [strain C57BL/6-TgN(ACTbEGFP)1Osb] (Okabe et al, 1997) were used to obtain NSCs constitutively expressing GFP as previously reported (Doncel-Pérez et al, 2009) (step 1). Neurospheres, isolated from E13 striatum, were incubated in NB27 medium (step 2), a mix of Neurobasal medium and B27 supplement (GIBCO, Scotland, UK) containing human bFGF (10 ng/mL) and EGF (20 ng/ml; Peprotech, New Jersey, USA), L-glutamine (0.5 mM), L-glutamate (25 mM), penicillin (100 U/mL), streptomycin (0.1 mg/mL) (all from Sigma-Aldrich), and fungizone (2.5 g/mL; Invitrogen, Madrid, Spain).…”

“…The therapeutic potential of transplantation of NSCs from transgenic embryos expressing green fluorescent protein (GFP) is assessed for lacunar stroke in this study. A previous study has characterized these cells by microarray analysis and in vitro neuritogenic activity as demonstrating a high potential for neural differentiation (Doncel-Pérez et al, 2009). We show that transplanted NSCs survived within the infarct area, and differentiated into neuronal and glial cells, specifically into striatal non-DA neurons and astrocytes.…”

Embryonic Neural Stem Cell Differentiation to Aldynoglia Induced by Olfactory Bulb Ensheathing Cell-Conditioned Medium

“…Initially, OBEC-conditioned medium was used to elicit neuritogenesis in vitro via the cocultures of neurons and EC (Kafitz & Greer, 1999;Sonigra et al, 1999;Wang et al, 2003), and more recently, we and other groups have been studying the capacity of the OBECconditioned medium to induce and direct in vitro differentiation from neural stem cells (Rojas-Mayorquín et al, 2008;Zhang et al, 2008;Doncel-Pérez et al, 2009) or even from other stem cells (Wang et al, 2007). Given that OBECs are able to produce, secrete and also respond to a variety of growth factors (Table 2) and because the OB displays constant neuroglial replacement (Altman, 1969;Graziadei and Monti Graziadei, 1978), it is therefore reasonable to propose that the OBEC could induce its own differentiation from stem cell precursors located in the OB in vivo, or even that of other stem cells in vitro (Rojas-Mayorquín et al, 2008;.…”