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首页> 外文期刊>Journal of Cellular Physiology >Midbrain cues dictate differentiation of human dental pulp stem cells towards functional dopaminergic neurons
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Midbrain cues dictate differentiation of human dental pulp stem cells towards functional dopaminergic neurons

机译：中脑线索决定分化的人类牙髓干细胞功能多巴胺神经元

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Dental pulp originating from the neural crest is considered a better source of postnatal stem cells for cell-based therapies in neurodegenerative diseases. Dental Pulp Stem Cells (DPSCs) have been shown to differentiate into cell-types of cranial neural crest ontology; however, their ability to differentiate to functional neurons of the central nervous system remains to be studied. We hypothesized that midbrain cues might commit DPSCs to differentiate to functional dopaminergic cell-type. As expected, DPSCs in their na?ve state spontaneously expressed early and mature neuronal markers like nestin, musashi12, β tubulin III, and Map2ab. On exposure to midbrain cues (sonic hedgehog, fibroblast growth factor 8 and basic fibroblast growth factor), DPSCs showed upregulation of dopaminergic neuron-specific transcription factors Nuclear Receptor related protein 1 (Nurr1), Engrailed 1 (En1) and paired-like homeodomain transcription factor 3 (Pitx3) as revealed by real-time RT-PCR. Immunofluorescence and flow cytometry analysis showed enhanced expression of mature neuronal marker Map2ab and dopaminergic-neuronal markers [tyrosine hydroxylase (TH), En1, Nurr1, and Pitx3], with nearly 77% of the induced DPSCs positive for TH. Functional studies indicated that the induced DPSCs could secrete dopamine constitutively and upon stimulation with potassium chloride (KCl) and adenosine triphosphate (ATP), as measured by dopamine ELISA. Additionally, the induced DPSCs showed intracellular Ca2+ influx in the presence of KCl, unlike control DPSCs. ATP-stimulated Ca2+ influx was observed in control and induced DPSCs, but only the induced cells secreted dopamine. Our data clearly demonstrate for the first time that DPSCs in the presence of embryonic midbrain cues show efficient propensity towards functional dopaminergic cell-type.
机译：牙髓起源于神经嵴认为是一个更好的产后干细胞来源细胞的细胞疗法神经退行性疾病。分化细胞(DPSCs)已被证明颅神经嵴细胞类型的本体;然而,他们的区分能力功能神经元的中枢神经系统还有待研究。中脑线索可能提交DPSCs区分多巴胺能程控功能。预期,DPSCs na的吗?早期和成熟的神经元自发地表达标记像巢蛋白、musashi12β微管蛋白三世,和Map2ab。刺猬,纤维母细胞生长因子8和基础纤维母细胞生长因子),DPSCs显示upregulation的多巴胺能特异性神经元转录因子核受体相关蛋白1 (Nurr1),十字的1 (En1)和paired-like homeodomain转录因子3实时rt - pcr (Pitx3)了。免疫荧光法和流式细胞术分析显示增强成熟神经元的表达标记Map2ab dopaminergic-neuronal标记酪氨酸羟化酶(TH)、En1 Nurr1,和Pitx3),近77%的诱导DPSCsTH阳性。,诱导DPSCs会分泌多巴胺既定的和刺激的氯化钾(氯化钾)和腺苷三磷酸腺苷(ATP),以多巴胺ELISA。细胞内钙离子涌入氯化钾的存在,与控制DPSCs。在控制和诱导DPSCs观察,但只有诱导细胞分泌多巴胺。数据明显首次展示DPSCs胚胎中脑的暗示显示有效的倾向于功能多巴胺能程控。

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