Innervating stackable neural organoid slices with tissue-like mesh electrodes for improved neural circuit development and characterization

神经类器官是研究人类大脑发育的极好工具，然而，它们并不能完全代表大脑结构。在神经类器官中长时间记录神经元活动也是困难的。为了应对这些挑战，在这个项目中，将开发一种新的方法来获得更好地模拟大脑结构的神经类器官。这些改进的类器官将被组装起来，以代表不同大脑区域之间的相互作用。该项目的第二个目标是将类似组织的超柔性网状电极整合到这些类器官中。已知这些网状电极对细胞的影响最小，并且可以安全地长期监测神经元的电活动。最后一个目标是研究对神经类器官施加电刺激是否可以加速它们的成熟，这通常需要几个月的时间。总之，该项目将导致改进的神经类器官模型和人类大脑发育的新知识。该项目还将支持一些教育和推广活动，包括新课程开发，本科生研究项目和带薪实习计划。 来自不同背景的积极性高的地区高中，本科生和研究生将有机会从事干细胞研究，这将有助于在这个令人兴奋的领域招募和留住学生。神经类器官为研究大脑发育和疾病提供了一个有前途的平台。然而，目前的神经类器官仍然受到缺乏适当的区域化和电刺激功能成熟和发育研究的慢性监测手段的限制。该项目旨在通过开发一种工程化的神经类器官系统来超越这些关键限制，该系统被适当地区域化并由能够长期监测和刺激的组织样网状系统支配，以改善对新皮层中电路发育的研究。总体研究目标将通过三个主要目标来实现：（1）通过堆叠切片的丘脑类器官和切片的端脑类器官与背腹图案化的皮层和腭下，组装能够实现丘脑-腭下-皮层投射的区域化神经类器官。(2)切片的类器官将由组织样的超柔性网状电极支配，以形成能够实时反馈慢性发育状态的切片类器官工程组装体（EASO）。(3)电刺激器将进一步集成到网状系统中，用于EASO的闭环调节和监测，促进加速发育研究的功能成熟。该项目将加深对神经回路发育的基本理解，促进类器官的发育。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

Neural organoids are excellent tools to study the development of human brains, however, they do not fully represent the brain structure. It is also difficult to record neuronal activities for a long time in neural organoids. To address those challenges, in this project, a new approach will be developed to derive neural organoids that better mimic brain structures. Those improved organoids will be assembled to represent interactions between various brain regions. The second objective of this project is to incorporate tissue-like, ultra-flexible mesh electrodes into those organoids. Those mesh electrodes are known to have minimal impacts on cells and can safely monitor the electrical activities of neurons chronically. The last objective is to investigate if applying electrical stimulations to neural organoids can accelerate their maturation, which usually takes several months. Together, this project will lead to improved neural organoid models and new knowledge in human brain development. The project will also support several educational and outreach activities, including new course development, undergraduate research projects, and paid internship programs. Highly motivated regional high school, undergraduate, and graduate students from diverse backgrounds will have opportunities to engage in stem cell research, which will facilitate the recruitment and retention of students in this exciting field.Neural organoids provide a promising platform for studying brain development and diseases. However, current neural organoids are still limited by the lack of proper regionalization and electrical stimulation for functional maturation and means of chronic monitoring for developmental studies. This project aims to transcend these key limitations by developing an engineered neural organoid system that is properly regionalized and innervated with a tissue-like mesh system capable of chronic monitoring and stimulation for improved studies of circuitry development in the neocortex. The overall research goal will be accomplished through three main objectives: (1) Regionalized neural organoid that enables the thalamus-subpallium-cortex projections will be assembled by stacking - sliced thalamic organoids and sliced telencephalic organoids with dorsoventrally patterned cortex and subpallium. (2) The sliced organoids will be innervated with tissue-like, ultra-flexible mesh electrodes for forming an Engineered Assembly of Sliced Organoids (EASOs) capable of real-time feedbacking the chronic developmental state. (3) Electrical stimulators will be further integrated into the mesh system for closed-loop modulation and monitoring of the EASOs, promoting functional maturation for accelerated developmental studies. The project will deepen the fundamental understanding of neural circuitry development and facilitate organoid development.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.