Development of forebrain organoid platform for modelling human cortical neurogenesis

开发用于模拟人类皮质神经发生的前脑类器官平台

基本信息

批准号：
9122502
负责人：
Guo-li Ming
金额：
$ 5.06万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9122502
关键词：
Address Adherens Junction Adherent Culture Affect Animal Model Apical Biological Process Bioreactors Brain Brain Diseases Brain Stem Cell Culture Techniques Cells Cerebral cortex Cerebrum Copy Number Polymorphism Culture Media DNA Sequence Alteration Development Disease Disease model Embryo Epilepsy Epileptogenesis Event Fibroblasts Forebrain Development Future Generalized Epilepsy Generations Glutamates Goals Growth Health Heterogeneity Human Human Development Human Engineering Incubators Intestines Investigation Kidney Laboratories Life Mental disorders Methodology Methods Midbrain structure Modeling Molecular Mus Names Nervous system structure Neuroepithelial Neuroglia Neurons Nutrient Organ Organogenesis Organoids Oxygen Patients Pluripotent Stem Cells Positioning Attribute Preclinical Drug Evaluation Predisposition Primates Production Property Prosencephalon Protocols documentation Radial Replacement Therapy Reporting Reproducibility Retinal Risk Rodent Role Sampling Schizophrenia Seizures Signal Transduction Skin Somatic Cell Stem Cell Research Stem cells Structure System Technology Testing Therapeutic Tissues Variant Work absorption base biological systems cell assembly cell type cellular development cost drug testing frontier genetic risk factor genome editing human disease improved in vivo induced pluripotent stem cell insight malformation microdeletion miniaturize mouse model nerve stem cell neurodevelopment neurogenesis novel novel strategies organ growth pluripotency prototype relating to nervous system self assembly stem cell differentiation synaptic function technology development three dimensional cell culture

项目摘要

DESCRIPTION (provided by applicant): A major breakthrough in the stem cell field over the last decade has been the development of technology to reprogram human somatic cells into induced pluripotent stem cells or iPSCs. In addition to the ability to derive specific cell types from human iPSCs in dish as 2D cultures, rapid progress in the field has made it possible to generate 3D cultures, or organoids, from iPSCs resembling whole developing organs, including intestinal, kidney, retinal, and cerebral cortex. Human organoids provide a unique opportunity to model organ development in a culture system that is similar to human organogenesis in vivo. Furthermore, organoid cultures provide the opportunity to model diseases that affect multiple cell types and to investigate non-cell- autonomous effects. The work in my laboratory focuses on neural development using both mouse models and iPSC models; and our long term goal is to understand mechanisms underlying normal brain development, neurodevelopmental diseases and to aid in the development of rational therapeutic strategies. Despite the tremendous promise of cerebral organoids to model brain genesis and brain diseases, there are several major limitations of the currently available technology, including high cost, low reproducibility and high variability, that limit our ability for quantitative analyses and broad application of the technology. We have recently developed a new approach by miniaturizing the critical components used to generate cerebral organoids, which allows for a dramatic reduction in materials, cell culture media, space and costs. In this exploratory project, we propose to further standardize forebrain specific organoid production and optimize cell culture conditions for directed and sustained growth. As a proof-of-principle, we will use this system to test the hypothesis that 15q11.2 microdeletion, a prominent genetic risk factor for epilepsy, leads to aberrant cortical neurogenesis for seizure susceptibility. We believe that our approach will transform organogenesis modeling and facilitate the identification of disease-relevant biological processes that are difficult to recapitulate in 2D monolayer cultures.

描述（由申请人提供）：过去十年干细胞领域的重大突破是除了能够从人类衍生特定细胞类型之外，还开发了将人类体细胞重编程为诱导性多能干细胞或 iPSC 的技术。培养皿中的 iPSC 作为 2D 培养物，该领域的快速进展使得从类似于整个发育器官的 iPSC 生成 3D 培养物或类器官成为可能，包括肠、肾、视网膜和人类类器官提供了在类似于体内人体器官发生的培养系统中模拟器官发育的独特机会，此外，类器官培养物还提供了模拟影响多种细胞类型的疾病和研究非细胞自主效应的机会。我实验室的工作重点是使用小鼠模型和 iPSC 模型进行神经发育；我们的长期目标是了解正常大脑发育、神经发育疾病的机制，并帮助制定合理的治疗策略，尽管脑神经发展前景广阔。模拟大脑起源和大脑的类器官疾病，目前可用技术存在几个主要局限性，包括成本高、重现性低和变异性大，限制了我们进行定量分析和广泛应用的能力我们最近开发了一种新方法，通过微型化用于生成大脑类器官的关键组件，可以大幅减少材料、细胞培养基、空间和成本。在这个探索性项目中，我们建议进一步标准化前脑特定类器官。作为原理验证，我们将使用该系统来测试 15q11.2 微缺失（一个重要的遗传风险因素）的假设。我们相信，我们的方法将改变器官发生模型，并有助于识别难以在 2D 单层培养物中重现的疾病相关生物过程。