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Khongkla Ekkaphot Uppakara Kwanchanok Boonmuen Nittaya Bhukhai Kanit Saengsawang Witchuda 《Cellular and molecular neurobiology》2022,42(7):2337-2353
Cellular and Molecular Neurobiology - The Cath.a-differentiated (CAD) cell line is a central nervous system-derived catecholaminergic cell line originating from tyrosine hydroxylase (TH)-producing... 相似文献
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Exosomes deliver functional proteins and genetic materials to neighboring cells, and have potential applications for tissue regeneration. One possible mechanism of exosome-promoted tissue regeneration is through the delivery of microRNA (miRNA). In this study, we hypothesized that exosomes derived from neuronal progenitor cells contain miRNAs that promote neuronal differentiation. We treated mesenchymal stem cells (MSCs) daily with exosomes derived from PC12 cells, a neuronal cell line, for 1 week. After the treatment with PC12-derived exosomes, MSCs developed neuron-like morphology, and gene and protein expressions of neuronal markers were upregulated. Microarray analysis showed that the expression of miR-125b, which is known to play a role in neuronal differentiation of stem cells, was much higher in PC12-derived exosomes than in exosomes from B16-F10 melanoma cells. These results suggest that the delivery of miRNAs contained in PC12-derived exosomes is a possible mechanism explaining the neuronal differentiation of MSC. 相似文献
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Dendritic cells have been investigated in clinical trials, predominantly with the aim of stimulating immune responses against tumours or infectious diseases. Thus far, however, no clinical studies have taken advantage of their specific immunosuppressive potential. Tolerogenic DCs may represent a new therapeutic strategy for human immune-based diseases, such as Crohn’s disease, where the perturbations of the finely tuned balance between the immune system and the microflora result in disease. In the present report, we describe the generation of tolerogenic DCs from healthy donors and Crohn’s disease patients using clinical-grade reagents in combination with dexamethasone as immunosuppressive agent and characterize their response to maturation stimuli. Interestingly, we found out that dexamethasone-conditioned DCs keep their tolerogenic properties to Gram-negative bacteria. Other findings included in this study demonstrate that the combination of dexamethasone with a specific cytokine cocktail yielded clinical-grade DCs with the following characteristics: a semi-mature phenotype, a pronounced shift towards anti-inflammatory versus inflammatory cytokine production and low T-cell stimulatory properties. Importantly, in regard to their clinical application, the tolerogenic phenotype of DCs remained stable after the elimination of dexamethasone and after a second stimulation with LPS or bacteria. All these properties make this cell product suitable to be tested in clinical trials of inflammatory conditions including Crohn’s disease. 相似文献
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Cecchi C Pensalfini A Liguri G Baglioni S Fiorillo C Guadagna S Zampagni M Formigli L Nosi D Stefani M 《Neurochemical research》2008,33(12):2516-2531
A substantial lack of information is recognized on the features underlying the variable susceptibility to amyloid aggregate
toxicity of cells with different phenotypes. Recently, we showed that different cell types are variously affected by early
aggregates of a prokaryotic hydrogenase domain (HypF-N). In the present study we investigated whether differentiation affects
cell susceptibility to amyloid injury using a human neurotypic SH-SY5Y cell differentiation model. We found that retinoic
acid-differentiated cells were significantly more resistant against Aβ1-40, Aβ1-42 and HypF-N prefibrillar aggregate toxicity
respect to undifferentiated cells treated similarly. Earlier and sharper increases in cytosolic Ca2+ and ROS with marked lipid peroxidation and mitochondrial dysfunction were also detected in exposed undifferentiated cells
resulting in apoptosis activation. The reduced vulnerability of differentiated cells matched a more efficient Ca2+-ATPase equipment and a higher total antioxidant capacity. Finally, increasing the content of membrane cholesterol resulted
in a remarkable reduction of vulnerability and ability to bind the aggregates in either undifferentiated and differentiated
cells.
Special issue article in honor of Dr. Anna Maria Giuffrida-Stella. 相似文献
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Embryonic Stem Cells (ESCs) represent an invaluable tool for the study of early mammalian development, for regenerative medicine
and for drug discovery. To fulfill these promises, efficient and easy protocols to differentiate ESCs have to be developed.
Most of these protocols results in low efficiency of neural induction and/or requires extended in vitro culture. Here we describe
in detail an easy and efficient method to differentiate ESCs into neurons, that can be used to identify molecules required
for proper neuronal differentiation. Moreover, we present a modification of this method that allows to clearly evaluate the
ability of some molecules to favor neuron formation in vitro. These methods can represent an efficient platform for studying
the molecular mechanisms underlying early events of neural induction and differentiation in ESCs, as well as for testing molecule
efficacy in the pharmaceutical testing. 相似文献
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Kaiyuan Wu Ruotong Ren Wenting Su Bo Wen Yuying Zhang Fei Yi Xinhua Qiao Tingting Yuan Jinhui Wang Limin Liu Juan Carlos Izpisua Belmonte Guang-Hui Liu Chang Chen 《The Journal of biological chemistry》2014,289(29):20193-20199
Alcohol dehydrogenase 5 (ADH5) is a conserved enzyme for alcohol and aldehyde metabolism in mammals. Despite dynamic expression throughout neurogenesis, its role in neuronal development remains unknown. Here we present the first evidence that ADH5 is a negative regulator of neuronal differentiation. Gene expression analyses identify a constant reduction of ADH5 levels throughout neuronal development. Overexpression of ADH5 reduces both development and adult neuronal differentiation of mouse neurons. This effect depends on the catalytic activity of ADH5 and involves ADH5-mediated denitrosation of histone deacetylase 2 (HDAC2). Our results indicate that ADH5 counteracts neuronal differentiation of human neural stem cells and that this effect can be reversed by pharmacological inhibition of ADH5. Based on these observations, we propose that ADH5 is a novel suppressor of neuronal differentiation and maturation. Inhibition of ADH5 may improve adult neurogenesis in a physiological or pathological setting. 相似文献
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Sebastian Weber María L. Fernández-Cachón Juliana M. Nascimento Steffen Knauer Barbara Offermann Robert F. Murphy Melanie Boerries Hauke Busch 《PloS one》2013,8(2)
Detection of neuronal cell differentiation is essential to study cell fate decisions under various stimuli and/or environmental conditions. Many tools exist that quantify differentiation by neurite length measurements of single cells. However, quantification of differentiation in whole cell populations remains elusive so far. Because such populations can consist of both proliferating and differentiating cells, the task to assess the overall differentiation status is not trivial and requires a high-throughput, fully automated approach to analyze sufficient data for a statistically significant discrimination to determine cell differentiation. We address the problem of detecting differentiation in a mixed population of proliferating and differentiating cells over time by supervised classification. Using nerve growth factor induced differentiation of PC12 cells, we monitor the changes in cell morphology over days by phase-contrast live-cell imaging. For general applicability, the classification procedure starts out with many features to identify those that maximize discrimination of differentiated and undifferentiated cells and to eliminate features sensitive to systematic measurement artifacts. The resulting image analysis determines the optimal post treatment day for training and achieves a near perfect classification of differentiation, which we confirmed in technically and biologically independent as well as differently designed experiments. Our approach allows to monitor neuronal cell populations repeatedly over days without any interference. It requires only an initial calibration and training step and is thereafter capable to discriminate further experiments. In conclusion, this enables long-term, large-scale studies of cell populations with minimized costs and efforts for detecting effects of external manipulation of neuronal cell differentiation. 相似文献
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目的:研究Tbx18是否能成功转染脂肪干细胞并使脂肪干细胞向心肌细胞分化。方法:分离培养来源于日本大耳兔腹股沟部脂肪的兔脂肪干细胞,用搭载有Tbx18的腺病毒载体转染脂肪干细胞,诱导分化后检测向心肌细胞的分化情况,同时将转染了含GFP的腺病毒组与未转染组作为对照。采用用流式细胞仪检测转染效率,采用免疫荧光法检测平滑肌肌动蛋白α-SMA,采用实时定量PCR法检测兔肌钙蛋白TNNT2的表达。结果:转染后荧光显微镜下可观察到荧光表达,且持续时间较长。流式细胞仪检测转染效率为70%左右;诱导分化后,脂肪干细胞内出现了α-SMA和TNNT2的表达。结论:Tbx18可成功转染入脂肪干细胞,且能在细胞内稳定表达;Tbx18可诱导脂肪干细胞向心肌样细胞分化。 相似文献
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It has been reported that rat bone marrow stromal cells (BMSCs) are differentiated into neuronal cells by administration of
2-mercaptoethanol [Woodbury et al (2000) J Neurosci Res 61:364–370]. In this study, we examined the effects of various sulfhydryl
(SH) compounds on the differentiation of BMSCs obtained from rat femurs. Neuronal differentiation was detected morphologically
and immunocytochemically. It was found that the cells treated with reduced glutathione (GSH) apparently differentiated into
neurons, showing extensive processes, and expressing neuron-specific enolase and microtubule-associated protein 2. Glutathione
monoethyl ester (GEE), which increased the cellular GSH content, showed no effect on the expression of neuronal markers. It
is concluded that the neural differentiation of BMSCs occurs by the administration of GSH. It was suggested that extracellular
and not intracellular GSH have effects on the induction of the neuronal differentiation of BMSCs. 相似文献
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Folic Acid Acts Through DNA Methyltransferases to Induce the Differentiation of Neural Stem Cells into Neurons 总被引:1,自引:0,他引:1
Suhui Luo Xumei Zhang Min Yu Hai Yan Huan Liu John X. Wilson Guowei Huang 《Cell biochemistry and biophysics》2013,66(3):559-566
The present study investigated the roles of folic acid and DNA methyltransferases (DNMTs) in the differentiation of neural stem cells (NSCs). Neonatal rat NSCs were grown in suspended neurosphere cultures and identified by their expression of SOX2 protein and capacity for self-renewal. Then NSCs were assigned to five treatment groups for cell differentiation: control (folic acid-free differentiation medium), low folic acid (8 μg/mL), high folic acid (32 μg/mL), low folic acid and DNMT inhibitor zebularine (8 μg/mL folic acid and 150 nmol/mL zebularine), and high folic acid and zebularine (32 μg/mL folic acid and 150 nmol/mL zebularine). After 6 days of cell differentiation, immunocytochemistry and western blot analyses were performed to identify neurons by β-tubulin III protein expression and astrocytes by GFAP expression. We observed that folic acid increased DNMT activity which may be regulated by the cellular S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), and the abundance of neurons but decreased the number of astrocytes. Zebularine blocked these effects of folic acid. In conclusion, folic acid acts through elevation of DNMT activity to increase neuronal differentiation and decrease astrocytic differentiation in NSCs. 相似文献
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Ras-Regulated Hypophosphorylation of the Retinoblastoma Protein Mediates Neuronal Differentiation in PC12 Cells 总被引:2,自引:0,他引:2
†HongZhen Li ‡Hiroshi Kawasaki ‡Eisuke Nishida Seisuke Hattori Shun Nakamura 《Journal of neurochemistry》1996,66(6):2287-2294
Abstract: To investigate the role of the retinoblastoma protein pRB in neuronal differentiation, we have measured the accumulation of hypophosphorylated pRB in PC12 cells stimulated by nerve growth factor (NGF). NGF induced the accumulation of hypophosphorylated pRB within 30 min and the level peaked after 12 h. Viral Kiras, cyclic AMP (cAMP), and 12- O -tetradecanoylphorbol 13-acetate (TPA) also induced the hypophosphorylation of pRB, but epidermal growth factor and interleukin-6 did not. The extent of hypophosphorylation of pRB correlated well with the capacity of these factors to stimulate neurite outgrowth. The constitutively activated Ras induced persistent shift of the phosphorylation state of pRB toward hypophosphorylation. A dominant negative form of cHa-Ras suppressed significantly induction of the hypophosphorylation of pRB by NGF, but not by cAMP. Taken together, these results suggest that the hypophosphorylation of pRB triggered by NGF is mediated by a Ras-dependent pathway. Furthermore, microinjection of a monoclonal antibody specific for the hypophosphorylated form of pRB blocked the neurite outgrowth initiated by NGF. These results suggest a crucial role of pRB in withdrawal of cells from the cell cycle and in neuronal differentiation of PC12 cells. 相似文献
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Pyramidal cells in the electrosensory lateral line lobe (ELL) of weakly electric fish have been observed to produce high-frequency burst discharge with constant depolarizing current (Turner et al., 1994). We present a two-compartment model of an ELL pyramidal cell that produces burst discharges similar to those seen in experiments. The burst mechanism involves a slowly changing interaction between the somatic and dendritic action potentials. Burst termination occurs when the trajectory of the system is reinjected in phase space near the ghost of a saddle-node bifurcation of fixed points. The burst trajectory reinjection is studied using quasi-static bifurcation theory, that shows a period doubling transition in the fast subsystem as the cause of burst termination. As the applied depolarization is increased, the model exhibits first resting, then tonic firing, and finally chaotic bursting behavior, in contrast with many other burst models. The transition between tonic firing and burst firing is due to a saddle-node bifurcation of limit cycles. Analysis of this bifurcation shows that the route to chaos in these neurons is type I intermittency, and we present experimental analysis of ELL pyramidal cell burst trains that support this model prediction. By varying parameters in a way that changes the positions of both saddle-node bifurcations in parameter space, we produce a wide gallery of burst patterns, which span a significant range of burst time scales. 相似文献
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《Developmental cell》2020,52(3):294-308.e3
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