Synthetic microfluidic synthesis of spinal cord tissues from human pluripotent stem cells

人类多能干细胞脊髓组织的微流体合成

• 批准号: 9805605
• 负责人: Jianping Fu
• 金额: $ 42.16万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9805605
• 关键词: 3-Dimensional; Address; Anatomy; Autologous; Biology; Cell Therapy; Cells; Chemical Stimulation; Chemicals; Code; Cyst; Development; Devices; Diagnosis; Disease model; Drug toxicity; Ectoderm; Embryo; Foundations; Gene Expression Profile; Generations; Genetic; Goals; Growth and Development function; Human; Impairment; Investigation; Lead; Life; Liquid substance; Measurement; Methodology; Microfluidic Microchips; Microfluidics; Modeling; Morphology; Nervous system structure; Neural Tube Development; Neural tube; Neuraxis; Neuroepithelial; Neuroepithelial Cells; Neurons; Organoids; Pathology; Pattern; Pattern Formation; Positioning Attribute; Prevention; Process; Property; Protocols documentation; Reproducibility; Research; Route; SHH gene; Signal Induction; Signal Transduction; Specific qualifier value; Spinal; Spinal Cord; Stem Cell Development; Stem cells; Structure; System; Target Populations; Tissues; Tubular formation; United States National Institutes of Health; base; cell transformation; human pluripotent stem cell; human tissue; innovation; innovative technologies; morphogens; nerve stem cell; nervous system development; nervous system disorder; neural patterning; neural plate; precursor cell; progenitor; programs; public health relevance; quantum; relating to nervous system; screening; self organization; smoothened signaling pathway; transcription factor

Project Summary During development of the vertebrate nervous system, a vast array of neurons will develop in discrete anatomical positions, acquire varied morphological forms, and establish connections with specific populations of target cells. Such spatial organization of cell fates and differentiation during the development of the nervous system are directed by concentration gradients of chemical signals, termed morphogens. Even though the importance of graded morphogen signaling in developmental pattern formation has been well recognized, it remains a significant question in biology about how embryonic progenitor cells transform dynamic changes in developmental signaling into spatial patterns of gene expression and cellular differentiation in a reliable and robust fashion. The long-term functional goal of this NIH R21 project is to specifically address the significant challenge in understanding the interpretation of morphogen gradients by intracellular signaling cascades while embryonic precursor cells are undergoing multicellular self-organization during developmental patterning. Specifically, we propose to leverage the intrinsic lumenogenic and self-organizing properties of neuroepithelial (NE) cells, the embryonic precursor cells in the neural tube, in conjunction with an innovative microfluidic embryological device, to achieve controllable and reproducible generations of lumenal NE cysts to mimic un- patterned spinal cord tissues. High-purity NE cells will be derived from human pluripotent stem cells (hPSCs) using established 2D directed differentiation protocols. Lumenal NE cysts will then be utilized seamlessly in the same microfluidic device for downstream asymmetrical patterning using the morphogen Sonic hedgehog (Shh) to achieve progressive acquisition of ventral neuronal subtypes in the spinal cord. Successful accomplishment of this proposed research will lead to the establishment of an innovative microfluidics-based methodology for controllable, reproducible, and scalable generation of (autologous) human spinal cord tissues from hPSCs, a quantum leap compared with existing 3D organoid culture systems that are known to lack controllability and reproducibility. Furthermore, our synthetic patterned human spinal cord model will provide a very useful experimental platform that offers superior experimental controls of key parameters and quantitative measurements to allow in-depth mechanistic investigations on the emergent self-organizing principles and pattering mechanisms that provide robustness and reliability to embryonic patterning, a long-standing question in biology.

项目摘要 在脊椎动物神经系统的开发过程中，会在离散中发展大量神经元 解剖位置，获得多样化的形态形式，并与特定人群建立联系 目标细胞。这种空间组织的细胞命运和神经发展过程中的分化 系统由化学信号的浓度梯度指导，称为形态学。即使 在发育模式形成中分级形态学信号传导的重要性已得到充分认识， 关于胚胎祖细胞如何改变动态变化的生物学仍然是一个重要的问题 在可靠和 强大的时尚。该NIH R21项目的长期功能目标是专门解决重要的问题 通过细胞内信号级联对形态梯度解释的挑战，而 胚胎前体细胞在发育模式期间正在经历多细胞自组织。 具体而言，我们提议利用神经上皮的内在腔源和自组织特性 （NE）细胞，神经管中的胚胎前体细胞，并与创新的微流体结合 胚胎设备，以实现可控和可重复的世代的腔内囊肿，以模仿 图案化的脊髓组织。高纯度NE细胞将源自人多能干细胞（HPSC） 使用已建立的2D定向分化协议。然后将无缝地使用Lumenal NE囊肿 使用形态学刺猬的同一用于下游不对称图案的微流体装置 （SHH）以逐步获得脊髓中腹神经元亚型的逐步获取。成功的 这项拟议的研究的完成将导致建立创新的基于微流体的研究 可控，可重现和可扩展生成（自体）人脊髓组织的方法 与现有的3D类器官培养系统相比，HPSC从HPSC中进行了量子飞跃 可控性和可重复性。此外，我们的合成图案化的人脊髓模型将提供 非常有用的实验平台，可为关键参数和定量提供卓越的实验控制 测量以允许对新兴自我组织原则和 具有对胚胎构图的稳健性和可靠性的图案机制，这是一个长期存在的问题 在生物学中。