Phenotypic & Functional Analysis of Human Schwann Cells for Potency Assay Dvlpmt

表型

基本信息

批准号：
8660106
负责人：
Paula Virginia Monje
金额：
$ 19万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8660106
关键词：
Address Adult Affect Afferent Neurons Animal Model Authorization documentation Autologous Axon Basal lamina Beds Biological Biological Assay Biological Markers Biology Biopsy Cell Culture Techniques Cell Therapy Cell Transplantation Cell Transplants Cell physiology Cell surface Cells Cellular Structures Characteristics Clinical Clinical Trials Coculture Techniques Cyclic AMP Data Development Differentiation Antigens FDA approved Goals Grant Growth Heterogeneity Human Human Activities In Vitro Injury Interleukin-2 Lesion Literature Measures Methods Mitogens Modification Myelin Natural regeneration Nerve Neurodegenerative Disorders Neurons Paralysed Patients Performance Peripheral Nerves Phase Phase I Clinical Trials Phenotype Population Property Protocols documentation Rattus Recovery of Function Relative (related person)Reporting Rodent Schwann Cells Signal Transduction Site Source Spinal cord injury Stimulus Study Subject Sulfoglycosphingolipids System Testing Therapeutic Transplantation Universities Work analytical method axon growth axon regeneration base cell type design evaluation/testing functional restoration human tissue improved in vitro Assay in vivo injured medical schools myelination novel public health relevance research clinical testing safety testing

项目摘要

DESCRIPTION (provided by applicant): The Schwann cell (SC) is a cell type of choice for cell transplantation therapies that aim to restore function in the injured CNS and PNS. Work in animal models has consistently shown that SCs reduce the size of the cavities that form after spinal cord injury; promote axon regeneration across the lesion site and myelinate regenerating axons, thereby improving functional recovery. In July 2012, the FDA approved a Phase I clinical trial to test the safety of autologous adult nerve-derived human SC transplantation in patients suffering from spinal cord injury. The feasibility to use primary SCs in cell therapy relies on th availability of cell culture methods rendering high numbers of transplantation quality purified SCs from peripheral nerve biopsies. An accurate assessment of the identity and function of the human SC product is essential to determine the potential therapeutic value of the cells. Indeed, the assessment of cell function prior to transplantation is a regulatory requirement for product release in later phases of clinical testing. Even though an extensive literature exists that report studies on rodent SCs, studies on human SCs are scarce. This is at least in part due to the poor performance of cultured human SCs in standard functional assays that were originally designed for rodent cells. This project seeks to overcome technical barriers in the development of in vitro assays to evaluate the biological activity of human SCs. Because SC identity and function are inextricably intertwined, we propose to begin addressing this challenge by developing simple analytical methods to evaluate the differentiating potential of SCs to become myelinating cells by the identification of differentiation markers before and after administration of cAMP, an instructing signal for differentiation (Aim 1). Because ensheathment and myelination of axons are perhaps the two most critical determinants for SC function in vitro and in vivo, we also propose to develop myelinating co-cultures of human SCs and neurons by implementing rational modifications to current protocols affecting both the SC and the neuronal components of the co-culture system (Aim 2). Identifying the most important cellular characteristics of in vitro expanded SCs relevant to their differentiating potential into myelin-forming cells will allow us to develop and implement reliable assays that assess the therapeutic value of the cells. If successful, co-cultures systems that allow for a quantification of myelin formation in vitro can serve as a direct measure of SC potency.

描述（由申请人提供）：Schwann细胞（SC）是一种细胞移植疗法的细胞类型，旨在恢复受伤的CNS和PNS中的功能。动物模型中的工作始终表明，SC会减少脊髓损伤后形成的空腔的大小；在病变部位促进轴突再生，并髓鞘再生轴突，从而改善功能恢复。 2012年7月，FDA批准了一项I期临床试验，以测试患有脊髓损伤患者的自体神经衍生的人类SC移植的安全性。在细胞疗法中使用原发性SC的可行性取决于细胞培养方法的可用性，从而使大量的移植质量纯化的SC来自周围神经活检。对人类SC产物的身份和功能的准确评估对于确定细胞的潜在治疗价值至关重要。实际上，移植前对细胞功能的评估是在临床测试后期阶段释放产品的调节要求。即使存在大量文献报道关于啮齿动物SC的研究，对人类SC的研究很少。这至少部分是由于最初为啮齿动物细胞设计的标准功能测定中培养的人类SC的性能不佳。该项目旨在克服体外测定的发展技术障碍，以评估人类SC的生物学活性。由于SC的身份和功能是密不可分的，因此我们建议通过开发简单的分析方法开始解决这一挑战，以评估SC的区分潜力，通过鉴定camp给药之前和之后的分化标记，这是一个指示信号的指示信号（AIM 1）。由于轴突的包裹和髓鞘化也许是体外和体内SC功能的两个最关键的决定因素，因此我们还建议通过对影响当前方案的合理修改来开发髓鞘的共培养，从而影响SC和COULTURE系统的神经元成分的当前方案（AIM AIM 2）。确定体外扩展的SC的最重要的细胞特征与它们与髓磷脂形成细胞的区分潜力相关的SC将使我们能够开发和实施可靠的测定方法，以评估细胞的治疗价值。如果成功，可以在体外量化髓磷脂形成的共同培养系统可以作为SC效力的直接衡量。