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Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies

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机构: [1]Cell Therapy Center, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, and Key Laboratory of Neurodegeneration, Ministry of Education [2]Center of Neural Injury and Repair, Beijing Institute for Brain Disorders
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DOI: 10.3791/59690
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关键词: Developmental Biology Issue 149 neuroscience induced neural stem cells Parkinson's disease reprogramming differentiation transplantation peripheral blood mononuclear cells Sendai virus 6-OHDA dopaminergic neurons

摘要:
Parkinson's disease (PD) is caused by degeneration of dopaminergic (DA) neurons at the substantia nigra pars compacta (SNpc) in the ventral mesencephalon (VM). Cell replacement therapy holds great promise for treatment of PD. Recently, induced neural stem cells (iNSCs) have emerged as a potential candidate for cell replacement therapy due to the reduced risk of tumor formation and the plasticity to give rise to region-specific neurons and glia. iNSCs can be reprogrammed from autologous somatic cellular sources, such as fibroblasts, peripheral blood mononuclear cells (PBMNCs) and various other types of cells. Compared with other types of somatic cells, PBMNCs are an appealing starter cell type because of the ease to access and expand in culture. Sendai virus (SeV), an RNA non-integrative virus, encoding reprogramming factors including human OCT3/4, SOX2, KLF4 and c-MYC, has a negative-sense, single-stranded, non-segmented genome that does not integrate into host genome, but only replicates in the cytoplasm of infected cells, offering an efficient and safe vehicle for reprogramming. In this study, we describe a protocol in which iNSCs are obtained by reprogramming PBMNCs, and differentiated into specialized VM DA neurons by a two-stage method. Then DA precursors are transplanted into unilaterally 6-hyroxydopamine (6-OHDA)-lesioned PD mouse models to evaluate the safety and efficacy for treatment of PD. This method provides a platform to investigate the functions and therapeutic effects of patient-specific DA neural cells in vitro and in vivo.

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出版当年[2018]版:
大类 | 4 区 综合性期刊
小类 | 4 区 综合性期刊
最新[2023]版:
大类 | 4 区 综合性期刊
小类 | 4 区 综合性期刊
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出版当年[2017]版:
Q2 MULTIDISCIPLINARY SCIENCES
最新[2023]版:
Q3 MULTIDISCIPLINARY SCIENCES

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第一作者:
第一作者机构: [1]Cell Therapy Center, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, and Key Laboratory of Neurodegeneration, Ministry of Education [2]Center of Neural Injury and Repair, Beijing Institute for Brain Disorders
通讯作者:
通讯机构: [1]Cell Therapy Center, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, and Key Laboratory of Neurodegeneration, Ministry of Education [2]Center of Neural Injury and Repair, Beijing Institute for Brain Disorders
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