Aalders Jeffrey, Léger Laurens, Tuerlings Tim, Ledda Sergio, van Hengel Jolanda
Medical Cell Biology research group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Corneel Heymanslaan 10, Building B, Entrance 36, 9000 Ghent, Belgium.
Department of Veterinary Medicine, University of Sassari, Sassari, Italy.
MethodsX. 2020 Sep 12;7:101065. doi: 10.1016/j.mex.2020.101065. eCollection 2020.
Three-dimensional (3D) cell culturing has several advantages over 2D cultures. 3D cell cultures more accurately mimic the environment, which is vital to obtain reliable results in disease modelling and toxicity testing. With the introduction of the Yamanaka factors, reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) became available. This iPSC technology provides a scalable source of differentiated cells. iPSCs can be programmed to differentiate into any cell type of the body, including cardiomyocytes. These heart-specific muscle cells, can then serve as a model for therapeutic drug screening or assay development. Current methods to achieve multicellular spheroids by 3D cell cultures, such as hanging drop and spinner flasks are expensive, time-consuming and require specialized materials and training. Hydrophobic powders can be used to create a micro environment for cell cultures, which are termed liquid marbles (LM). In this procedure we describe the first use of the LM technology for 3D culturing derived human cardiomyocytes which results in the formation of cardiospheres within 24h. The cardiospheres could be used for several in depth and high-throughput analyses.
与二维细胞培养相比，三维（3D）细胞培养具有多个优势。3D细胞培养能更准确地模拟环境，这对于在疾病建模和毒性测试中获得可靠结果至关重要。随着山中因子的引入，将体细胞重编程为诱导多能干细胞（iPSC）成为可能。这种iPSC技术提供了一种可扩展的分化细胞来源。iPSC可被编程分化为体内任何细胞类型，包括心肌细胞。这些心脏特异性肌肉细胞随后可作为治疗性药物筛选或检测方法开发的模型。目前通过3D细胞培养实现多细胞球体的方法，如悬滴法和转瓶培养法，成本高、耗时，且需要特殊材料和培训。疏水性粉末可用于为细胞培养创造微环境，这种微环境被称为液滴弹（LM）。在本实验中，我们描述了首次将LM技术用于3D培养人源心肌细胞，该技术可在24小时内形成心脏球体。这些心脏球体可用于多种深入和高通量分析。
