用于研究脑类器官的实验和计算方法:综述

Experimental and Computational Methods for the Study of Cerebral Organoids: A Review.

作者信息

Poli Daniele, Magliaro Chiara, Ahluwalia Arti

机构信息

Research Center E. Piaggio, University of Pisa, Pisa, Italy.

Department of Information Engineering, University of Pisa, Pisa, Italy.

出版信息

Front Neurosci. 2019 Mar 5;13:162. doi: 10.3389/fnins.2019.00162. eCollection 2019.

Abstract

Cerebral (or brain) organoids derived from human cells have enormous potential as physiologically relevant downscaled models of the human brain. In fact, these stem cell-derived neural aggregates resemble the three-dimensional (3D) cytoarchitectural arrangement of the brain overcoming not only the unrealistic somatic flatness but also the planar neuritic outgrowth of the two-dimensional (2D) cultures. Despite the growing use of cerebral organoids in scientific research, a more critical evaluation of their reliability and reproducibility in terms of cellular diversity, mature traits, and neuronal dynamics is still required. Specifically, a quantitative framework for generating and investigating these models of the human brain is lacking. To this end, the aim of this review is to inspire new computational and technology driven ideas for methodological improvements and novel applications of brain organoids. After an overview of the organoid generation protocols described in the literature, we review the computational models employed to assess their formation, organization and resource uptake. The experimental approaches currently provided to structurally and functionally characterize brain organoid networks for studying single neuron morphology and their connections at cellular and sub-cellular resolution are also discussed. Well-established techniques based on current/voltage clamp, optogenetics, calcium imaging, and Micro-Electrode Arrays (MEAs) are proposed for monitoring intra- and extra-cellular responses underlying neuronal dynamics and functional connections. Finally, we consider critical aspects of the established procedures and the physiological limitations of these models, suggesting how a complement of engineering tools could improve the current approaches and their applications.

摘要

源自人类细胞的大脑类器官作为与人类大脑生理相关的缩小模型具有巨大潜力。事实上,这些干细胞衍生的神经聚集体类似于大脑的三维(3D)细胞结构排列,不仅克服了不切实际的体细胞扁平状态,还克服了二维(2D)培养中神经突的平面生长。尽管大脑类器官在科学研究中的应用越来越广泛,但仍需要从细胞多样性、成熟特征和神经元动力学方面对其可靠性和可重复性进行更严格的评估。具体而言,缺乏用于生成和研究这些人类大脑模型的定量框架。为此,本综述的目的是激发新的计算和技术驱动的想法,以改进大脑类器官的方法并实现其新应用。在概述文献中描述的类器官生成方案后,我们回顾了用于评估其形成、组织和资源摄取的计算模型。还讨论了目前用于在细胞和亚细胞分辨率下对大脑类器官网络进行结构和功能表征以研究单个神经元形态及其连接的实验方法。提出了基于电流/电压钳、光遗传学、钙成像和微电极阵列(MEA)的成熟技术,用于监测神经元动力学和功能连接背后的细胞内和细胞外反应。最后,我们考虑了既定程序的关键方面以及这些模型的生理局限性,提出了工程工具如何补充可以改进当前方法及其应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ed4/6411764/205911f750c2/fnins-13-00162-g001.jpg

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