Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies
机构:[1]Cell Therapy Center, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, and Key Laboratory of Neurodegeneration, Ministry of Education内科系统首都医科大学宣武医院细胞生物学研究室[2]Center of Neural Injury and Repair, Beijing Institute for Brain Disorders
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.
基金:
Stem Cell and Translation National Key Project [2016YFA0101403]; National Natural Science Foundation of China [81661130160, 81422014, 81561138004]; Beijing Municipal Natural Science Foundation [5142005]; Beijing Talents Foundation [2017000021223TD03]; Support Project of High-level Teachers in Beijing Municipal Universities in the Period of 13th Five-year Plan [CIT TCD20180333]; Beijing Medical System High Level Talent Award [2015-3-063]; Beijing Municipal Health Commission Fund [PXM 2018_026283_000002]; Beijing One Hundred, Thousand, and Ten Thousand Talents Fund [2018A03]; Beijing Municipal Administration of Hospitals Clinical Medicine Development of Special Funding Support [ZYLX201706]; Royal Society-Newton Advanced Fellowship [NA150482]
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
推荐引用方式(GB/T 7714):
Zheng Wei,Chen Zhiguo.Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies[J].JOVE-JOURNAL OF VISUALIZED EXPERIMENTS.2019,(149):-.doi:10.3791/59690.
APA:
Zheng, Wei&Chen, Zhiguo.(2019).Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies.JOVE-JOURNAL OF VISUALIZED EXPERIMENTS,,(149)
MLA:
Zheng, Wei,et al."Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies".JOVE-JOURNAL OF VISUALIZED EXPERIMENTS ..149(2019):-