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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要在脊椎动物神经系统的发育过程中，大量的神经元会以离散的方式发育。解剖位置，获得不同的形态形式，并与特定人群建立联系的靶细胞。神经发育过程中细胞命运和分化的这种空间组织系统由化学信号（称为形态发生素）的浓度梯度引导。尽管分级形态发生素信号传导在发育模式形成中的重要性已得到充分认识，它关于胚胎祖细胞如何转化动态变化仍然是生物学中的一个重要问题发育信号以可靠且可靠的方式转化为基因表达和细胞分化的空间模式强劲时尚。 NIH R21 项目的长期功能目标是专门解决重要的问题理解细胞内信号级联对形态发生素梯度的解释是一个挑战，同时胚胎前体细胞在发育模式过程中经历多细胞自组织。具体来说，我们建议利用神经上皮的内在发光和自组织特性 (NE) 细胞，神经管中的胚胎前体细胞，与创新的微流体相结合胚胎学装置，以实现管腔 NE 囊肿的可控和可重复世代，以模仿非有图案的脊髓组织。高纯度 NE 细胞将源自人类多能干细胞 (hPSC) 使用已建立的 2D 定向分化方案。然后，腔内 NE 囊肿将被无缝地用于使用相同的微流体装置，使用形态发生素 Sonic Hedgehog 进行下游不对称图案化（嘘）以实现脊髓中腹侧神经元亚型的逐步获取。成功的这项拟议研究的完成将导致建立一个基于微流体的创新可控、可重复和可扩展的（自体）人类脊髓组织生成方法与已知缺乏的现有 3D 类器官培养系统相比，这是一个巨大的飞跃可控性和再现性。此外，我们的合成图案人类脊髓模型将提供非常有用的实验平台，可提供关键参数和定量的卓越实验控制测量以允许对新兴的自组织原理进行深入的机械研究为胚胎模式提供鲁棒性和可靠性的模式机制，这是一个长期存在的问题在生物学中。