Induced Pluripotent Stem Cells from Neurosurgical removed tissue for Brain Repair

描述（由申请人提供）：该项目的长期目标是从诱导多能干细胞（iPSC）技术中开发患者特异性自体细胞来源，作为创伤性脑损伤和其他神经系统疾病的治疗方法。目前许多神经移植方法都遇到了与供体细胞来源相关的问题。神经移植作为一种治疗手段应用于临床之前，需要克服的突出问题包括：种族争议、细胞来源有限、移植后存活率低、移植细胞功能缺乏整合、体内肿瘤形成和免疫排斥反应等。因此，确定更有利于细胞替代疗法的最佳细胞来源至关重要。人类iPSC生成的新兴技术，随后定向分化成均匀的神经祖细胞（hNP）和神经元群体，作为对从TBI患者获得的人类细胞进行逆向工程的方法，具有很大的前景。在这个项目中，我们的工作重点是对从神经外科切除组织中分离出来的成年人神经细胞进行重编程。该提议的具体假设是，功能性iPSC衍生的神经祖细胞可以从神经外科切除组织中产生，并用作TBI的细胞替代疗法。该假设是基于我们的观察和研究进展，即（a）iPSC可以从患有特定神经退行性疾病的患者产生，（B）多能干细胞的定向分化可以产生均匀的hNP群体，和（c）成人干细胞样细胞可以从神经外科切除组织中分离并成功培养为神经球和单层。为了验证这一假设，在本提案中，我们将首先从神经外科切除组织中分离的成人神经细胞中产生iPSCs，并评估其多能特性，体外定向分化能力。然后，我们将评估这些iPSC衍生的hNP在完整免疫缺陷动物和受损免疫活性动物的脑中的体内移植潜力。提出的两个具体目标是：（1）从神经外科切除组织中分离的成人神经细胞生成和表征iPSC。(2)评估iPSC衍生的NPs在受损环境中的存活、分化和功能，并评估性别差异对细胞存活的作用。所提出的研究的结果将对开发患者特异性hiPSC作为TBI和其他神经系统疾病的自体细胞替代策略产生重大影响。

DESCRIPTION (provided by applicant): The long-term goal of this project is to develop patient-specific autologous cell source from induced pluripotent stem cells (iPSC) technology as a therapy for traumatic brain injury and other neurological diseases. Many current neural transplantation approaches have encountered problems associated with the source of donor cells. The ethnic controversies, limited cell availability, poor survival, lack of functional integration of grafted cells, the risk of tumor formation and immunological rejection in vivo are prominent issues need to be overcome before neural transplantation as a therapy to be used in clinic. It is essential, therefore, to identify optimal cell sources that are more conducive for cel replacement therapies. The emerging technology of human iPSCs generation, followed by directed differentiation into uniform populations of neural progenitor cells (hNPs) and neurons, holds great promise as an approach to reverse engineer human cells obtained from TBI patients. In this project, we are focusing our efforts on reprogramming adult human neural cells isolated from neurosurgical resection tissues. The specific hypothesis of this proposal is that functional iPSC-derived neural progenitor cells can be generated from neurosurgical resection tissues and utilized as cell replacement therapy for TBI. The hypothesis is based on our observations and research developments that (a) iPSCs can be generated from patients with specific neurodegenerative disorders, (b) directed differentiation of pluripotent stem cells can lead to uniform populations of hNPs, and (c) adult human stem-like cells can be isolated from neurosurgical resection tissues and successfully cultured as neurospheres and monolayers. To test the hypothesis, in this proposal, we will first generate iPSCs from adult human neural cells isolated from neurosurgical resection tissues and evaluate their pluripotent characteristics, directed differentiation capabilities in vitro. We will then assess the transplantation potential o these iPSC-derived hNPs in vivo in the brain in both intact immunodeficient animals and injured immunocompetent animals. The two proposed specific aims are: (1) Generate and characterize iPSCs from adult human neural cells isolated from neurosurgical resection tissues. (2) Evaluate the survival, differentiation and functionality of iPSC-derived NPs in the injured environment and assess the role of gender differences on cell survival. Results from the proposed study will have significant impact in the development of patient-specific hiPSCs as autologous cell replacement strategies for TBI and other neurological disorders.