Zuppinger Christian
Cardiology Department, DBMR MEM C812, University Hospital Bern, Bern, Switzerland.
Methods Mol Biol. 2019;1929:41-52. doi: 10.1007/978-1-4939-9030-6_4.
There is a need for organotypic in vitro models that resemble the native tissue in functionality and tissue architecture for disease models and drug development. To this end, many 3D culture formats have been developed over time. Among the most often used type is the scaffold-free multicellular aggregate, also called spheroid, that forms by self-assembly. However, working with 3D cultures can be challenging because single cells are not as accessible as in 2D cultures and standard lab procedures must be adapted or replaced altogether. This chapter describes methods to create cardiac spheroids consisting of human iPSC-derived cardiomyocytes and cardiac fibroblasts and how to measure contractility or calcium signals using quantitative video analysis and confocal microscopy. Emphasis is on the particular challenges that 3D cultures pose and on affordable methods that do not require specialized equipment.
对于疾病模型和药物开发而言，需要在功能和组织结构上类似于天然组织的器官型体外模型。为此，随着时间的推移，已经开发出许多三维培养形式。最常用的类型之一是无支架多细胞聚集体，也称为球体，它通过自组装形成。然而，使用三维培养可能具有挑战性，因为单个细胞不像二维培养那样易于操作，并且标准实验室程序必须进行调整或完全替换。本章描述了创建由人诱导多能干细胞衍生的心肌细胞和心脏成纤维细胞组成的心脏球体的方法，以及如何使用定量视频分析和共聚焦显微镜测量收缩性或钙信号。重点是三维培养带来的特殊挑战以及不需要专门设备的经济实惠的方法。
