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

描述（由申请人提供）：雪旺细胞（SC）是一种用于细胞移植治疗的细胞类型，旨在恢复受损中枢神经系统和PNS的功能。动物模型的研究一致表明，SCs减少了脊髓损伤后形成的空腔的大小；促进整个损伤部位的轴突再生和髓鞘再生轴突，从而改善功能恢复。2012年7月，FDA批准了一项I期临床试验，以测试自体成人神经源性人SC移植在脊髓损伤患者中的安全性。在细胞治疗中使用原代SCs的可行性依赖于细胞培养方法的可用性，从周围神经活检中获得大量移植质量的纯化SCs。准确评估人类SC产物的特性和功能对于确定细胞的潜在治疗价值至关重要。事实上，在移植前对细胞功能的评估是临床试验后期产品释放的监管要求。尽管有大量的文献报道

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.