Development of a validated human iPSCs-derived microglia and brain organoid co-culture platform to investigate neuro-immune interactions

该项目的总体目标是开发一个经过验证的人类小胶质细胞和皮质脑类器官共培养平台，以研究神经免疫相互作用。人脑类器官是来自培养皿中生长的人类多能干细胞（hPSC）的脑细胞的3D自组装结构。脑类器官模拟人类大脑发育的关键方面，这使它们成为研究大脑发育和神经系统疾病的人类特定方面的优秀模型系统。然而，脑类器官天生只产生外胚层来源的脑细胞，而不是来自其他谱系的细胞，如小胶质细胞。小胶质细胞是脑中的常驻免疫细胞，占人脑中总细胞的约5-10%，并且在病理条件下在脑发育、稳态和神经炎症中起重要作用。因此，建立hPSC衍生的小胶质细胞整合到3D人脑类器官中的平台为研究人类小胶质细胞和模拟脑疾病提供了独特的机会。以前的研究表明，产生含有小胶质细胞的类器官的可行性未能验证该系统。在这个项目中，我们的目标是优化hPSC衍生的小胶质细胞和脑类器官共培养方法，通过系统比较不同的条件和新的方法，并通过比较它与新鲜的人胚胎组织在细胞，转录组和功能水平的平台进行验证。此外，我们将开发一个管道，用于从转录组数据中对神经元-免疫相互作用进行生物信息学分析。

The overarching goal of this project is to develop a validated human microglia and cortical brain organoid co-culture platform to investigate neuro-immune interactions. Human brain organoids are 3D self-assembled structures of brain cells derived from human pluripotent stem cells (hPSCs) that are grown in a dish. Brain organoids mimic key aspects of human brain development, which makes them an excellent model system for studying human-specific aspects of brain development and neurological disorders. However, brain organoids innately generate only ectodermal-originated brain cells not cells from other lineages such as microglia. Microglia are resident immune cells in the brain comprising about 5-10% of total cells in the human brain and play important roles in brain development, homeostasis, and neuroinflammation during pathological conditions. Therefore, establishing a platform for hPSC-derived microglia integrated into 3D human brain organoids provides unique opportunities to study human microglia and to model brain disorders. Previous studies that have shown the feasibility of generating microglia-containing organoids failed to validate the system. In this project, we aim to optimize hPSC-derived microglia and brain organoid co-culture method by systematic comparisons of different conditions and novel approaches and validate the platform by comparing it with fresh human embryonic tissue at the cellular, transcriptomic, and functional levels. Furthermore, we will develop a pipeline for bioinformatic analyses of neuron-immune interactions from transcriptomic data.