Spheroid Formation and Evaluation of Hepatic Cells in a Three-Dimensional Culture Device
Spheroid Formation and Evaluation of Hepatic Cells in a Three-Dimensional Culture Device
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DOI：
10.3727/215517915x689056
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发表时间：
2015-08-01
期刊：
影响因子：
--
通讯作者：
Hayashi, Shuji
中科院分区：
文献类型：
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作者：
Miyamoto, Yoshitaka;Ikeuchi, Masashi;Hayashi, Shuji
在药物发现中，评估生物体内的肝细胞非常重要。与2D培养方法相比，肝细胞3D培养技术的发展成功地维持了长期的肝脏功能，形成了肝脏特异性结构。开展药物试验的关键是建立稳定的体外评价体系。在这篇文章中，我们报告了一个锥形支架集群培养（TASCL）设备开发，以创建肝脏球体在体外。TASCL装置将用作药物发现的毒性评价系统。TASCL装置的总尺寸为10 mm × 10 mm，包含4个微孔，每个微孔具有顶部孔（500 μ m × 500 μ m）和底部孔（300 μ m直径圆形）。我们评估了TASCL装置中HepG 2球状体的形成、恢复和大小。HepG 2球状体的形成和回收均接近100%，并且随着初始细胞接种密度的增加，球状体的大小增加。在微孔中球状体的大小没有显著差异。此外，使用TASCL装置获得的HepG 2球状体是活的并产生白蛋白。使用FE-SEM研究HepG 2球状体的形态。微孔中的球状体表现出完美的球形聚集。在本报告中，通过调整TASCL装置微孔的大小，创建了均匀的HepG 2球体，该装置有助于更精确地测量每个HepG 2球体的肝功能。我们的TASCL设备将适用于作为药物测试的毒性评估系统。
In drug discovery, it is very important to evaluate liver cells within an organism. Compared to 2D culture methods, the development of 3D culture techniques for liver cells has been successful in maintaining long-term liver functionality with the formation of a hepatic-specific structure. The key to performing drug testing is the establishment of a stable in vitro evaluation system. In this article, we report a Tapered Stencil for Cluster Culture (TASCL) device developed to create liver spheroids in vitro. The TASCL device will be applied as a toxicity evaluation system for drug discovery. The TASCL device was created with an overall size of 10 mm x 10 mm, containing 4(X) microwells with a top aperture (500 mu m x 500 mu m) and a bottom aperture (300 mu m diameter circular) per microwell. We evaluated the formation, recovery, and size of HepG2 spheroids in the TASCL device. The formation and recovery were both nearly 100%, and the size of the HepG2 spheroids increased with an increase in the initial cell seeding density. There were no significant differences in the sizes of the spheroids among the microwells. In addition, the HepG2 spheroids obtained using the TASCL device were alive and produced albumin. The morphology of the HepG2 spheroids was investigated using FE-SEM. The spheroids in the microwells exhibited perfectly spherical aggregation. In this report, by adjusting the size of the microwells of the TASCL, device, uniform HepG2 spheroids were created, and the device facilitated more precise measurements of the liver function per HepG2 spheroid. Our TASCL device will be useful for application as a toxicity evaluation system for drug testing.