Changes in HepG2 spheroid behavior induced by differences in the gap distance between spheroids in a micropatterned culture system
Changes in HepG2 spheroid behavior induced by differences in the gap distance between spheroids in a micropatterned culture system
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DOI：
10.1016/j.jbiosc.2017.12.013
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发表时间：
2018-06-01
影响因子：
2.8
通讯作者：
Nakazawa, Kohji
中科院分区：
文献类型：
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作者：
Miyamoto, Daisuke;Hara, Takuya;Nakazawa, Kohji
微图案化是一种很有前途的调控培养环境的技术。在这项研究中，我们研究了球体分离距离对它们的性质在HepG2球体的微图案化芯片的效果。基本的芯片设计包括37个胶原蛋白斑点（直径300 μ m）在玻璃基板上的六边形排列;没有胶原蛋白斑点的区域被聚乙二醇修饰以产生非粘性表面。制造了三个类似的芯片，胶原蛋白点之间的间隙距离为500，1000和1500 μ m。HepG2细胞贴附于胶原斑点上，并通过细胞增殖形成球状体。尽管HepG2球状体的白蛋白分泌活性在所有芯片中几乎相同，但当差距距离小于1000 μ m时，球状体生长和无氧代谢的抑制增强。此外，这种现象是由球体之间的干涉效应引起的，在芯片的内部区域比外部区域更明显。但当差距大于1500 μ m时，球体间的干涉效应几乎可以避免。此外，溶解氧的浓度在相邻的球体之间的差距距离的减少而减少，这表明球体竞争氧，并成为缺氧的方式，取决于球体的分离距离。这些结果表明，球体间距是一个重要的因素，可以调节球体的性质。(C)2017年，日本生物技术学会。All rights reserved.
Micropatterning is a promising technique for modulating culture environments. In this study, we investigated the effect of spheroid separation distance on their properties in a micropatterned chip of HepG2 spheroids. The basic chip design consisted of 37 collagen spots (300 mu m in diameter) in a hexagonal arrangement on a glass substrate; the region without collagen-spots was modified by polyethylene glycol to create the non-adhesive surface. Three similar chips were fabricated with gap distances between collagen-spots of 500, 1000, and 1500 mu m. HepG2 cells adhered on the collagen spots and then formed spheroids via cell proliferation. Although the albumin secretion activities of HepG2 spheroids were almost the same in all chips, inhibition of spheroid growth and anaerobic metabolism were intensified when the gap distance was less than 1000 mu m. Additionally, such phenomena which are induced by interference effects between spheroids, were more pronounced at the inside region of the chip than at the outside region. However, the interference effect between spheroids was nearly avoided when the gap distance was at least 1500 mu m. Furthermore, the concentration of dissolved oxygen between neighboring spheroids decreased as the gap distance decreased, indicating that the spheroids competed for oxygen and became hypoxic in a way that depended on the spheroid separation distance. These results indicate that the spheroid separation distance is an important factor that can modulate the spheroid properties. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.