KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 02841, Republic of Korea.
Biofabrication. 2017 Nov 30;10(1):015001. doi: 10.1088/1758-5090/aa9876.
The engineered three-dimensional (3D) cell cultivation system for the production of multicellular spheroids has attracted considerable attention due to its improved in vivo relevance to cellular communications compared with the traditional two-dimensional (2D) cell culture platform. The formation and maintenance of cell spheroids in a healthy condition is the critical factor for tissue engineering applications such as the repair of damaged tissues, the development of organ replacement parts and preclinical drug tests. However, culturing spheroids in conventional isolated single wells shows limited yield and reduced maintenance periods due to the lack of proper supplies of nutrition as well as intercellular chemical signaling. Here, we develop novel networked concave microwell arrays for the effective construction of 3D multi-cellular spheroids. The proposed method provides a suitable structure for the diffusion of oxygen, water-soluble nutrients and cytokines for cell-cell interactions between the spheroids in neighboring microwells. We have further demonstrated that hepatocyte spheroid cultured networked concave microwells show enhanced cell viability and albumin secretion compared to the un-networked control group over two weeks. Our results reveal that multi-cellular functionality can be tuned up by networking individual 3D spheroids without supplying additional chemicals or biological supplements. We anticipate our result to be useful in high-throughput cellular screening platforms to study cell-cell interactions, in response to diverse chemical stimuli as well as the development of the in vivo mimicking of the customized 3D tissue culture system.
由于与传统的二维（2D）细胞培养平台相比，用于生产多细胞球体的工程化三维（3D）细胞培养系统在细胞通信方面具有更高的体内相关性，因此引起了广泛关注。健康状态下细胞球体的形成和维持是组织工程应用的关键因素，例如受损组织的修复、器官替代部件的开发和临床前药物测试。然而，由于缺乏适当的营养供应以及细胞间化学信号传递，在传统的单个孤立孔中培养球体，其产量有限且维持时间缩短。在这里，我们开发了用于有效构建 3D 多细胞球体的新型网络凹形微井阵列。该方法为相邻微井中球体之间的氧气、水溶性营养物和细胞因子的扩散提供了合适的结构，从而促进了细胞-细胞相互作用。我们进一步证明，与未联网的对照组相比，在两周的时间内，肝细胞球体在网络凹形微井中的培养显示出更高的细胞活力和白蛋白分泌。我们的结果表明，通过将单个 3D 球体联网，可以在不添加其他化学物质或生物添加剂的情况下增强多细胞功能。我们预计我们的结果将有助于高通量细胞筛选平台，以研究细胞-细胞相互作用，以及针对各种化学刺激以及定制 3D 组织培养系统的体内模拟的发展。
