Synthetic substrates for the expansion and differentiation of hPSC-derived NPCs

项目摘要 尽管治疗进展，神经退行性疾病和障碍仍然是一些主要原因 美国的死亡率和发病率。因此，以细胞为基础的疗法，以取代失去或损坏 神经元或支持神经细胞具有很大的治疗意义。人神经前体细胞 来源于人多能干细胞（hPSC，包括人胚胎干细胞[hESC]和人 诱导多能干细胞[hiPSC]）可以广泛增殖并分化成所有神经谱系 (i.e.神经元、星形胶质细胞和少突胶质细胞），其危害中枢神经系统（CNS）。 因此，hNPCs及其分化的后代可以提供细胞原材料来模拟或治疗 各种神经系统疾病。然而，这些细胞的临床应用将需要（i）定义， 用于其扩增和神经元分化的无异种的条件，和（ii）可扩展的培养系统， 使它们能够扩增和神经元分化，数量足以用于再生医学和药物治疗。 筛选的目的。为此，我们将使用一种新的高通量方法来系统地筛选一种 合理设计的物理化学定义的聚合物库，以鉴定用于 hNPCs的扩增和神经元分化。接下来，我们将以这些合成底物为基础 用于工程化基于低剪切生物反应器的系统，用于大规模hNPC扩增和神经元扩增。 分化将评估按比例放大的细胞群的异质性以及它们的细胞， 生物化学、遗传学和电生理学特性。这项研究的成功完成将 显著推进hNPC及其衍生物的临床应用，因为它将使大规模的 以疾病建模，药物筛选， 和再生医学的应用。

PROJECT SUMMARY ABSTRACT Despite therapeutic advances, neurodegenerative diseases and disorders remain some of the leading causes of mortality and morbidity in the United States. Therefore, cell-based therapies to replace lost or damaged neurons or supporting neural cells are of great therapeutic interest. Human neural progenitor cells (hNPCs) derived from human pluripotent stem cells (hPSCs, including human embryonic stem cells [hESCs] and human induced pluripotent stem cells [hiPSCs]) can proliferate extensively and differentiate into all the neural lineages (i.e. neurons, astrocytes, and oligodendrocytes) that compromise the central nervous system (CNS). Therefore, hNPCs and their differentiated progeny could provide the cellular raw material to model or treat a variety of nervous system disorders. However, the clinical application of these cells will require (i) defined, xeno-free conditions for their expansion and neuronal differentiation and (ii) scalable culture systems that enable their expansion and neuronal differentiation in numbers sufficient for regenerative medicine and drug screening purposes. To that end, we will use a novel high-throughput approach to systematically screen a rationally designed library of physicochemically-defined polymers to identify candidate synthetic substrates for the expansion and neuronal differentiation of hNPCs. Next, we will use these synthetic substrates as the basis for the engineering of low shear bioreactor-based systems for large-scale hNPC expansion and neuronal differentiation. The scaled-up cell populations will be assessed for their heterogeneity as well as their cellular, biochemical, genetic, and electrophysiological properties. The successful completion of this research will significantly advance the clinical application of hNPCs and their derivatives as it will enable the large-scale expansion and neuronal differentiation of hNPCs in quantities necessary for disease modeling, drug screening, and regenerative medicine applications.