Medical Cell Biology research group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium.
Department of Veterinary Medicine, University of Sassari, Sassari, Italy.
Methods Mol Biol. 2021;2273:85-102. doi: 10.1007/978-1-0716-1246-0_5.
Cells have a remarkable ability to self-organize and rearrange in functional organoids, this process was greatly boosted by the recent advances in 3D culture technologies and materials. Presently, this approach can be applied to model human organ development and function "in a dish" and can be used to predict drug response in a patient specific fashion.Here we describe a protocol that allows for the derivation of functional cardiac mini organoids consisting of cocultured cardiomyocytes and cardiac fibroblast. Cells are suspended in a drop of medium and encapsulated with hydrophobic fumed silica powder nanoparticles. These nanoparticles are treated with hydrophobic chemicals, hexamethyldisilazane (nHMDS), and result in the formation of microbioreactors. These microenvironments are defined as "liquid marbles," stimulating cell coalescence and 3D aggregation. Then nHMDS shell ensures optimal gas exchange between the interior liquid and the surrounding environment. This microbioreactor makes working in smaller volumes possible and is therefore amenable for higher throughput applications. Moreover, the properties of liquid marble microbioreactors makes it an excellent culture technique for cocultures. Here we demonstrate how cocultures of cardiac fibroblast and cardiomyocytes in a cardiosphere can be a valuable tool to model cardiac diseases in vitro and to assess cell interactions to decipher disease mechanisms.
细胞具有自我组织和在功能性类器官中重新排列的非凡能力，这一过程得益于 3D 培养技术和材料的最新进展。目前，这种方法可用于在“培养皿”中模拟人类器官的发育和功能，并且可以用于以患者特异性的方式预测药物反应。在这里，我们描述了一种允许衍生功能性心脏微类器官的方案，该方案由共培养的心肌细胞和心脏成纤维细胞组成。细胞悬浮在培养基滴中，并被疏水性气相二氧化硅纳米粉末颗粒包封。这些纳米颗粒用疏水性化学品六甲基二硅氮烷（nHMDS）处理，导致形成微生物反应器。这些微环境被定义为“液体大理石”，刺激细胞聚合并进行 3D 聚集。然后，nHMDS 壳确保内部液体和周围环境之间的最佳气体交换。这种微生物反应器使在较小体积下工作成为可能，因此适用于高通量应用。此外，液体大理石微生物反应器的特性使其成为共培养的绝佳培养技术。在这里，我们展示了如何在心脏球体中培养心脏成纤维细胞和心肌细胞的共培养物，这是一种在体外模拟心脏疾病和评估细胞相互作用以破译疾病机制的有价值的工具。
