17ß-Oestradiol promotes differentiation of human embryonic stem cells into dopamine neurons via cross-talk between insulin-like growth factors-1 and oestrogen receptor ß.

Human embryonic stem cells (hESCs) can self-renew and differentiate into all cell lineages. E2 is known to exhibit positive effects on embryo development. Although the importance of E2 in many physiological processes has been reported, to date few researchers have investigated the effects of E2 on hESCs differentiation. We studied the effects of E2 on dopamine (DA) neuron induction of hESCs and its related signalling pathways using the three-stage protocol. In our study, 0.1 µM E2 were applied to hESCs-derived human embryoid bodies (hEBs) and effects of E2 on neural cells differentiation were investigated. Protein and mRNA level assay indicated that E2 up-regulated the expression of insulin-like growth factors (IGF)-1, ectoderm, neural precursor cells (NPC) and DA neuron markers, respectively. The population of hESC-derived NPCs and DA neurons was increased to 92% and 93% to that of DMSO group, respectively. Furthermore, yield of DA neuron-secreted tyrosine hydroxylase (TH) and dopamine was also increased. E2-caused promotion was relieved in single inhibitor (ICI or JB1) group partly, and E2 effects were repressed more stronger in inhibitors combination (ICI plus JB1) group than in single inhibitor group at hEBs, hNPCs and hDA neurons stages. Owing to oestrogen receptors regulate multiple brain functions, when single or two inhibitors were used to treat neural differentiation stage, we found that oestrogen receptor (ER)ß but not ERα is strongly repressed at the hNPCs and hDA neurons stage. These findings, for the first time, demonstrate the molecular cascade and related cell biology events involved in E2-improved hNPC and hDA neuron differentiation through cross-talk between IGF-1 and ERß in vitro.

Neuronal Cell Differentiation of Human Dental Pulp Stem Cells on Synthetic Polymeric Surfaces Coated With ECM Proteins.

Stem cells serve as an ideal source of tissue regeneration therapy because of their high stemness properties and regenerative activities. Mesenchymal stem cells (MSCs) are considered an excellent source of stem cell therapy because MSCs can be easily obtained without ethical concern and can differentiate into most types of cells in the human body. We prepared cell culture materials combined with synthetic polymeric materials of poly-N-isopropylacrylamide-co-butyl acrylate (PN) and extracellular matrix proteins to investigate the effect of cell culture biomaterials on the differentiation of dental pulp stem cells (DPSCs) into neuronal cells. The DPSCs cultured on poly-L-ornithine (PLO)-coated (TPS-PLO) plates and PLO and PN-coated (TPS-PLO-PN) plates showed excellent neuronal marker (ßIII-tubulin and nestin) expression and the highest expansion rate among the culture plates investigated in this study. This result suggests that the TPS-PLO and TPS-PN-PLO plates maintained stable DPSCs proliferation and had good capabilities of differentiating into neuronal cells. TPS-PLO and TPS-PN-PLO plates may have high potentials as cell culture biomaterials for the differentiation of MSCs into several neural cells, such as cells in the central nervous system, retinal cells, retinal organoids and oligodendrocytes, which will expand the sources of cells for stem cell therapies in the future.

Focus on 1,25-Dihydroxyvitamin D3 in the Peripheral Nervous System.

In this review, we draw attention to the roles of calcitriol (1,25-dihydroxyvitamin D3) in the trophicity of the peripheral nervous system. Calcitriol has long been known to be crucial in phosphocalcium homeostasis. However, recent discoveries concerning its involvement in the immune system, anti-cancer defenses, and central nervous system development suggest a more pleiotropic role than previously thought. Several studies have highlighted the impact of calcitriol deficiency as a promoting factor of various central neurological diseases, such as multiple sclerosis, amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease. Based on these findings and recent publications, a greater role for calcitriol may be envisioned in the peripheral nervous system. Indeed, calcitriol is involved in myelination, axonal homogeneity of peripheral nerves, and neuronal-cell differentiation. This may have useful clinical consequences, as calcitriol supplementation may be a simple means to avoid the onset and/or development of peripheral nervous-system disorders.

Resveratrol can inhibit Notch signaling pathway to improve spinal cord injury.

Spinal cord injury (SCI) is one of the severe central nervous system (CNS) diseases. Although various therapeutic approaches have been researched, there are still no effective therapeutic measures for SCI. Development of novel, safe and effective therapeutics for SCI have currently gained a lot of interest. Medicinal and chemical agents have the tendency of regulating signaling pathways. Notch signaling pathway plays an important role in neuronal cell differentiation, neuroinflammation and axonal regeneration in the wake of SCI. Resveratrol has been reported to exert neuroprotective effects in CNS conditions. Resveratrol can inhibit Notch signaling pathway to curtail SCI. Herein, we systematically review potential mechanisms of resveratrol-inhibiting Notch signaling pathway in SCI treatment.

Recombinant Spider Silk and Collagen-Based Nerve Guidance Conduits Support Neuronal Cell Differentiation and Functionality in Vitro.

Biomaterial scaffolds are under investigation as therapeutic tools to bridge nerve endings following traumatic peripheral nerve injury. The goal is to develop biocompatible nerve guidance conduits (NGCs) with internal guiding structures that promote longitudinally oriented cell migration and regeneration. In the present study, a nonwoven mesh (NWM) made of a recombinant spider silk protein was processed into a tubular structure, ensuring structural integrity of enclosed microfluidics-produced collagen fibers for cell and neurite guidance. The differentiated type of the neuroblastoma X glioma hybrid cell line NG108-15 was used as a model for studying neuronal differentiation on the individual components and on the complete NGC. Differentiated NG108-15 cells grown on recombinant spider silk NWM and collagen fibers formed neuronal networks and synapses. Additionally, whole-cell patch clamp recordings confirmed that all components supported the differentiation of NG108-15 cells into functional neurons. Our NGC demonstrated that tubes made of recombinant spider silk NWM filled with microfluidics-produced collagen fibers are well suited for peripheral nerve repair.

Neuronal Cell Morphology in Primary Cerebellar Granule Cells Using High-Content Analysis.

Neurite outgrowth, one of the underlying cellular processes that defines the development and functionality of the mammalian nervous system, is also a sensitive indicator of neuronal cell health. From screening libraries of putative neurotherapeutic compounds to analyzing the millions of environmental pollutants for which we have inadequate neurotoxicity safety data, the large volume of chemical compounds that require evaluation is a major obstacle for manual imaging and analysis methods. In this context, high-content analysis (HCA) has emerged as a sensitive and accurate method for detecting changes in neuronal cell morphology within a format applicable to screening large chemical libraries. Advances in HCA technologies have enabled the automated imaging and quantitative analysis of neurite outgrowth morphology within a 96-well plate in less than 5 min. Traditionally, neurite outgrowth assessment has been conducted on immortalized cell lines such as pheochromocytoma (PC-12) cells that differentiate into neuron-like cells upon culture with nerve growth factor. Unfortunately, they do not retain all the in vivo characteristics of physiological neuronal tissue, including lack of synapse formation. As researchers refine neurite outgrowth quantitative analysis using HCA, an emerging question is how to quantify this biology in more complex models that more faithfully recapitulate in vivo environments. Primary neurons provide several benefits relative to neuronal cell lines, including the elaboration of axons from secondary dendrites and formation of both pre- and postsynaptic junctions. This chapter reviews techniques for evaluating neurite outgrowth using the ArrayScan HCA platform within a model system of primary cultures of rodent cerebellar granule cells.

Bisphenol A Represses Dopaminergic Neuron Differentiation from Human Embryonic Stem Cells through Downregulating the Expression of Insulin-like Growth Factor 1.

Bisphenol A (BPA) is a ubiquitous compound emerging as a possible toxicant during embryonic development. Human embryonic stem cell (hESC) promises a valuable model for evaluating the effects of environmental chemicals on human prenatal development. In our study, 1 µM BPA were applied to hESC-derived embryoid bodies (hEBs) and effects of BPA on neural cell differentiation were investigated. The expression level of insulin-like growth factor 1 (IGF-1) and marker genes for ectoderm, neuron progenitor cells, and dopaminergic (DA) neurons were all repressed upon BPA exposure. The population of hESC-derived neural precursor cells (NPCs) and DA neurons were decreased. Furthermore, yield of DA neuron-secreted tyrosine hydroxylase (TH) and dopamine were also reduced. When recombinant IGF-1 supplied, BPA-caused repressions were partially or completely relieved. Our further methylation microarray analysis indicated that there was a higher methylation level on the promoter of SRY-related HMG-box 5 (SOX5), a possible enhancer of IGF-1. Consistently, next quantitative polymerase chain reaction (qPCR) results confirmed that SOX5 expression was downregulated. Our investigation suggests that BPA represses DA neuron differentiation mainly through downregulating IGF-1 expression, which may attribute to the altered methylation level on the promoter of IGF-1 upstream genes. Our findings first elaborate the mechanism of IGF-1-mediated BPA effects on neuronal differentiation, which is helpful to illuminate the unique mechanism of BPA toxicity on prenatal neurodevelopment.