Stem Cell-Seeded ECM Scaffolds for Neurotransplantation

描述(申请人提供)：神经移植在受损的中枢神经系统(CNS)中的成功有限。据推测，这种恶劣的环境可能需要通过定向细胞外基质(ECM)蛋白质工程来改善细胞支持，以改善移植物存活和细胞功能，从而促进功能恢复。本研究项目的总体目标是开发微创移植技术，通过基于ECM的支架优化干细胞/祖细胞的附着，并利用相关的实验模型(包括体外和体内)进行优化和结果评估。这一总体目标分为3个相互关联的具体目标：(1)鉴定基于神经干细胞(NS)细胞的3-D构建物在体外的特性，以实现微创移植策略和最大细胞存活率，并诱导期望的程度的增殖、迁移和分化；(2)通过在体外恶劣的替代环境中测试NS-ECM构建物，确定构建物整合的机制；(3)通过将构建体移植到挫伤小鼠脑内并检测损伤后宿主挫伤、细胞行为、认知和感觉运动行为结果的变化，分析组织工程构建物的体内功能。这项研究具有重要意义，因为它为祖细胞/干细胞移植技术提供了一种新的方法，并对结果进行了详细分析。这项研究可能会直接应用于神经外科的临床实践，从而允许治疗性的细胞替代治疗创伤性脑和脊髓损伤以及中枢神经系统退行性疾病。此外，这项研究将有助于深入了解中枢神经系统再生的机制，并有助于阐明神经移植成功所必需的细胞环境。通过分析控制良好的多水平系统的结果，这项研究还可能导致去细胞移植方法学的建立，并建立非胚胎/胎儿细胞来源移植所必需的要求。

DESCRIPTION (provided by applicant): Neural transplantation in the injured central nervous system (CNS) has had limited success. It is hypothesized that this hostile environment may require improved cell support through directed extracellular matrix (ECM) protein engineering to improve graft survival and cell function and hence, functional recovery. The overall objective of this research project is to develop minimally-invasive transplantation techniques for optimizing stem/progenitor cell attachment through ECM-based scaffolds and to utilize relevant experimental models (both in vitro and in vivo) for optimization and outcome assessment. This overall goal is divided into 3 interrelated specific aims: (1) To characterize neural stem (NS) cell-ECM-based 3-D constructs in vitro for minimally invasive grafting strategies and maximal cell survival and to elicit a desired degree of proliferation, migration, and differentiation; (2) To determine mechanisms of construct integration by testing NS-ECM constructs in a surrogate hostile in vitro environment; and (3) To analyze the in vivo function of tissue-engineered constructs by transplanting constructs into contused mouse brains and examining post-injury alterations in the host contusion, cell behavior, and cognitive and sensorimotor behavioral outcome. The research proposed is significant because it offers a novel approach to progenitor/stem cell transplant technology with detailed analyses of outcome. This research may have direct application to clinical practice in neurosurgery that would permit therapeutic, cellular replacement in the treatment of traumatic brain and spinal cord injuries and degenerative diseases of the CNS. In addition, this research will provide insight into the mechanisms of CNS regeneration and help to elucidate the necessary cellular environment for neurotransplantation success. By analyzing outcome in well-controlled multi-level systems, this research may also lead to acellular transplantation methodology and establish the requirements necessary for the transplantation of non-embryonic/fetal cell sources.