使用传统和创新方法从 iPSC 来源的牙髓干细胞生成和表征皮质类器官。

Generation and characterization of cortical organoids from iPSC-derived dental pulp stem cells using traditional and innovative approaches.

机构信息

Hospital Israelita Albert Einstein, São Paulo, SP, Brazil.

Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.

出版信息

Neurochem Int. 2024 Nov;180:105854. doi: 10.1016/j.neuint.2024.105854. Epub 2024 Sep 4.

Abstract

Cortical organoids derived from human induced pluripotent stem cells (hiPSCs) represent a powerful in vitro experimental system to investigate human brain development and disease, often inaccessible to direct experimentation. However, despite steady progress in organoid technology, several limitations remain, including high cost and variability, use of hiPSCs derived from tissues harvested invasively, unexplored three-dimensional (3D) structural features and neuronal connectivity. Here, using a cost-effective and reproducible protocol as well as conventional two-dimensional (2D) immunostaining, we show that cortical organoids generated from hiPSCs obtained by reprogramming stem cells from human exfoliated deciduous teeth (SHED) recapitulate key aspects of human corticogenesis, such as polarized organization of neural progenitor zones with the presence of outer radial glial stem cells, and differentiation of superficial- and deep-layer cortical neurons and glial cells. We also show that 3D bioprinting and magnetic resonance imaging of intact cortical organoids are alternative and complementary approaches to unravel critical features of the 3D architecture of organoids. Finally, extracellular electrical recordings in whole organoids showed functional neuronal networks. Together, our findings suggest that SHED-derived cortical organoids constitute an attractive model of human neurodevelopment, and support the notion that a combination of 2D and 3D techniques to analyze organoid structure and function may help improve this promising technology.

摘要

源自人类诱导多能干细胞(hiPSC)的皮质类器官代表了一种强大的体外实验系统,可用于研究人类大脑发育和疾病,而这些在直接实验中往往无法实现。然而,尽管类器官技术取得了稳步进展,但仍存在一些局限性,包括成本高、可变性大、使用的 hiPSC 来源于侵入性采集的组织、尚未探索的三维(3D)结构特征和神经元连接。在这里,我们使用具有成本效益且可重复的方案以及传统的二维(2D)免疫染色,证明了从人脱落乳牙(SHED)重编程的干细胞获得的 hiPSC 产生的皮质类器官再现了人类皮质发生的关键方面,例如具有外放射状胶质干细胞的神经祖细胞区的极化组织,以及浅层和深层皮质神经元和神经胶质细胞的分化。我们还表明,完整皮质类器官的 3D 生物打印和磁共振成像是揭示类器官 3D 结构关键特征的替代和互补方法。最后,整个类器官的细胞外电记录显示出功能性神经网络。总之,我们的研究结果表明,SHED 衍生的皮质类器官构成了人类神经发育的有吸引力的模型,并支持这样一种观点,即结合 2D 和 3D 技术来分析类器官的结构和功能可能有助于改进这项有前途的技术。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索