Selective stimulatory action of olfactory ensheathing glia-conditioned medium on oligodendroglial differentiation, with additional reference to signaling mechanisms

Abstract

We examined the effects of conditioned medium from olfactory ensheathing glia (OEGCM) on the differentiation of oligodendrocytes in mixed cultures of early postnatal hippocampi. Differentiation was judged from the numerical density (ND) of cells immunoreactive to 2'3' cyclic nucleotide 3'phosphodiesterase (CNPase) and O4 antibodies. NDs increased according to inverted-U dose-response curves, particularly for CNPase+ cells (9-fold at optimal dilution) and these changes were blocked by inhibitors of ERK1, p38-MAPK, and PI3K. Our results raise the possibility that OEG secreted factor(s) may counteract demyelination induced by trauma, neurodegenerative diseases, and advanced age, and should stimulate novel methods to deliver these factors and/or potentiating chemicals.

Streptococcus pneumoniae resists intracellular killing by olfactory ensheathing cells but not by microglia

“…as modified by our group720. Eight-week-old male Wistar rats were obtained from the Center for Experimental Surgery Laboratory at the Federal University of Rio de Janeiro.…”

Activation of cannabinoid type 1 receptor (CB1) modulates oligodendroglial process branching complexity in rat hippocampal cultures stimulated by olfactory ensheathing glia-conditioned medium

“…OEGs primary cell cultures were harvested from the olfactory bulb (OB) of adult Wistar rats (3-4 months) as previously reported (Carvalho et al, 2014), and at least 20 animals were used in this study. The OEG primary cell culture was also performed following a protocol previously described (Nash et al, 2001;Carvalho et al, 2014). Briefly, the olfactory fiber layer from the OB was dissociated and purified by differential adhesion (Nash et al, 2001), and then cells were placed on coverslips, T25 plates and/or 6-well plates previously treated with laminin (40 µg/ml, Sigma Aldrich) in complete DMEM-F12 supplemented with 10% fetal bovine serum and incubated in 5% CO 2 at 37 • C. After the OEG cell culture reached 80% confluence, the OEGCM was collected, filtered (0.22 µm membrane pore), aliquoted and stored at −70 • C until use.…”

“…We first isolated and cultured olfactory ensheathing glia (OEG) from the olfactory bulbs of adult rats following a protocol previously established by us and others (Nash et al, 2001;Carvalho et al, 2014Carvalho et al, , 2018. We also investigated if OEG in culture expresses components of the ECS, i.e., cannabinoid receptor type 1 (CB1), and the main endocannabinoid degradation enzymes, fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), which degrade anandamide (AEA) and 2-arachidonoyl glycerol (2-AG), respectively, by qRT-PCR (Figure 1A).…”

Olfactory Ensheathing Cells: A Trojan Horse for Glioma Gene Therapy

“…Olfactory ensheathing cells were purified from the olfactory bulbs of C57BL/6 male adult mice following our and others' previously described protocol (11)(12)(13)(14)(15). Briefly, the mice were killed by isoflurane.…”

“…We purified the olfactory ensheathing cells from the olfactory bulbs of C57BL/6 male adult mice following previously established protocol (11)(12)(13)(14)(15) (summarized in Figure 1A). Costaining of OECs for 2 0 ,3 0 cyclic nucleotide 3 0 -phosphodiesterase, smooth muscle a-actin, glial fibrillary acidic protein, and calciumbinding protein b, specific markers for OEC (13), revealed that our cultures displayed neither unlabeled cells nor cells labeled with only one marker, confirming that our method is highly effective for OEC purification (Figure 1B).…”