Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media
Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media
复制标题
DOI：
10.1016/j.jfma.2014.09.003
复制
发表时间：
2014-12-01
影响因子：
3.2
通讯作者：
Lin, Chun-Pin
中科院分区：
文献类型：
--
作者：
Chang, Chia-Chieh;Chang, Kai-Chun;Lin, Chun-Pin
背景/目的：牙髓干细胞（DPSC）由于其接近胚胎起源和易于获取，已被认为是神经再生干细胞的一个有前途的来源。本研究的目的是评估多巴胺能和运动神经元诱导培养基对人DPSCs（hDPSCs）向神经元样细胞转分化的影响。用XTT [（2，3-双-（2-甲氧基-4-硝基-5-磺基苯基）-2H-四唑鎓-5-甲酰苯胺）]测定评价DPSC的增殖潜力。制备用于诱导多巴胺能和脊髓运动神经元分化的培养基。通过免疫细胞化学和实时定量逆转录聚合酶链反应（RT-PCR）检测神经细胞特异性细胞标志物在DPSCs中的表达来评估神经诱导的效果。结果：在XTT试验中，在多巴胺能或运动神经元诱导培养基中培养的DPSCs分别减少2.6倍或2倍。与在对照培养基中培养的DPSC相比，在两种神经元诱导培养基中培养的DPSC中β III-微管蛋白（β III-tub）、胶质细胞酸性蛋白（GFAP）和少突胶质细胞（O 1）阳性细胞的比例显著较高。此外，hDPSC衍生的多巴胺能和脊髓运动神经元细胞诱导后表达更高密度的神经元细胞标志物比那些前induction.Conclusion：这些研究结果表明，在响应神经元诱导刺激，更大比例的DPSC停止增殖，并获得类似成熟神经元的表型。这种神经嵴衍生的成体DPSC可以为神经元疾病和损伤的基于疗法的治疗提供替代的干细胞来源。版权所有（C）2014，爱思唯尔台湾有限公司及台湾医学会. All rights reserved.
Background/Purpose: Dental pulp stem cells (DPSCs) have been proposed as a promising source of stem cells in nerve regeneration due to their close embryonic origin and ease of harvest. The aim of this study was to evaluate the efficacy of dopaminergic and motor neuronal inductive media on transdifferentiation of human DPSCs (hDPSCs) into neuron-like cells.Methods: Isolation, cultivation, and identification of hDPSCs were performed with morphological analyses and flow cytometry. The proliferation potential of DPSCs was evaluated with an XTT [(2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)] assay. Media for the induction of dopaminergic and spinal motor neuronal differentiation were prepared. The efficacy of neural induction was evaluated by detecting the expression of neuron cell-specific cell markers in DPSCs by immunocytochemistry and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR).Results: In the XTT assay, there was a 2.6-or 2-fold decrease in DPSCs cultured in dopaminergic or motor neuronal inductive media, respectively. The proportions of beta III-tubulin (beta III-tub), glial fibrillary acidic protein (GFAP), and oligodendrocyte (O1)-positive cells were significantly higher in DPSCs cultured in both neuronal inductive media compared with those cultured in control media. Furthermore, hDPSC-derived dopaminergic and spinal motor neuron cells after induction expressed a higher density of neuron cell markers than those before induction.Conclusion: These findings suggest that in response to the neuronal inductive stimuli, a greater proportion of DPSCs stop proliferation and acquire a phenotype resembling mature neurons. Such neural crest- derived adult DPSCs may provide an alternative stem cell source for therapy- based treatments of neuronal disorders and injury. Copyright (C) 2014, Elsevier Taiwan LLC & Formosan Medical Association. All rights reserved.