Effects of selegiline, a monoamine oxidase B inhibitor, on differentiation of P19 embryonal carcinoma stem cells, into neuron-like cells

Abstract

Selegiline, the irreversible inhibitor of monoamine oxidase B (MAO-B), is currently used to treat Parkinson's disease. However, the mechanism of action of selegiline is complex and cannot be explained solely by its MAO-B inhibitory action. It stimulates gene expression, as well as expression of a number of mRNAs or proteins in nerve and glial cells. Direct neuroprotective and anti-apoptotic actions of selegiline have previously been observed in vitro. Previous studies showed that selegiline can induce neuronal phenotype in cultured bone marrow stem cells and embryonic stem cells. Embryonal carcinoma (EC) cells are developmentally pluripotene cells which can be differentiated into all cell types under the appropriate conditions. The present study was carried out to examine the effects of selegiline on undifferentiated P19 EC cells. The results showed that selegiline treatment had a dramatic effect on neuronal morphology. It induced the differentiation of EC cells into neuron-like cells in a concentration-dependent manner. The peak response was in a dose of selegiline significantly lower than required for MAO-B inhibition. The differentiated cells were immunoreactive for neuron-specific proteins, synaptophysin, and β-III tubulin. Stem cell therapy has been considered as an ideal option for the treatment of neurodegenerative diseases. Generation of neurons from stem cells could serve as a source for potential cell therapy. This study suggests the potential use of combined selegiline and stem cell therapy to improve deficits in neurodegenerative diseases.

Differentiation of P19 embryonal carcinoma stem cells into insulin-producing cells promoted by pancreas-conditioned medium

“…They can be considered as complementary tools to study Table S1 the developmental mechanisms. The P19 mouse EC line is an excellent and widely used model to analyse regulation of neuronal development and differentiation (Bakhshalizadeh et al, 2011;Esmaeili et al, 2006;MacPherson and McBurney, 1995). P19 cells have a normal male karyotype (McBurney and Rogers, 1982).…”

The combined effects of three-dimensional cell culture and natural tissue extract on neural differentiation of P19 embryonal carcinoma stem cells

“…The dye does not stain glial cells and labels differentially neuronal from glial cells (Peinado et al, ). Light microscopy sections stained by cresyl violet technique have been previously used for cell counting and morphometric analysis (Bakhshalizadeh et al, ; Esmaeili et al, ; Peinado et al, ; Portiansky et al, ). Figure shows the morphology of the differentiated cells in 2D (exposed to different concentrations of NRBE) and 3D (cultured on BG − and BG + scaffolds) culture systems using cresyl violet.…”

“…The staining solution (0.25% cresyl violet, 0.8% glacial acetic acid, 0.6‐mM sodium acetate) was applied to the cells (Fraichard et al, ). As a preliminary evaluation of neuronal induction, five microscopic fields were randomly selected to count the stained cells (i.e., neuronal phenotype cells) and percentage of neuronal induction (PNI) was determined (Bakhshalizadeh, Esmaeili, Houshmand, Shirzad, & Saedi, ; Esmaeili, Tiraihi, Movahedin, & Mowla, ; Portiansky, Nishida, Barbeito, Gimeno, & Goya, ). The cells cultured with no tissue extract and/or scaffold were counted and considered as negative controls.…”

Induction of Tyrosine Hydroxylase Gene Expression in Embryonal Carcinoma Stem Cells Using a Natural Tissue‐Specific Inducer

“…The drug has antioxidant, antiapoptotic, antiaging, and neuroprotective effects (Knoll, ; Bakhshalizadeh et al , ; Miklya, ). In addition, deprenyl is able to reduce neurotoxin secretion and induce neural differentiation of stem cells and expression of a number of genes including nerve growth factor (NGF), brain‐derived neurotrophic factor (BDNF), neurotrophin‐3 (NT‐3), β‐III tubulin, and synaptophysin (Esmaeili et al , ; Bakhshalizadeh et al , ). Both deprenyl and BDNF increase the total length of the TH‐positive neurites (Iwasaki et al , ).…”