Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array
Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array
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DOI：
10.1002/biot.201300559
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发表时间：
2014-07-01
影响因子：
4.7
通讯作者：
Nakazawa, Kohji
中科院分区：
文献类型：
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作者：
Sakai, Yusuke;Hattori, Koji;Nakazawa, Kohji
微流体装置允许对三维 (3D) 组织进行灌注培养，模拟我们体内血管化 3D 组织中的血液流动。在这里，我们报告了一种微流体装置，由两部分微流体室芯片和多微孔阵列芯片组成，能够在培养终点拆卸。在微流体室内，可以形成 3D 组织聚集体（球体）阵列并在灌注下进行培养。随后，可以对从拆卸的装置收集的球体进行详细的培养后分析。该装置有助于均匀的球体形成、高通量格式的生长分析、通过灌注流速控制增殖以及球体的培养后分析。我们使用该装置在两种受控灌注流速下培养人肝细胞癌细胞 (HepG2) 细胞球体。 HepG2球体在较高灌注流速下比在较低灌注流速下表现出更大的细胞生长，并且在不同流速条件下表现出不同的代谢活性以及mRNA和蛋白质表达。这些结果显示了灌注培养在精确控制微流体装置中的培养环境方面的潜力。球体阵列室的构造允许同时测试多种培养条件，在毒性和药物筛选方面具有潜在的应用。
Microfluidic devices permit perfusion culture of three-dimensional (3D) tissue, mimicking the flow of blood in vascularized 3D tissue in our body. Here, we report a microfluidic device composed of a two-part microfluidic chamber chip and multi-microwell array chip able to be disassembled at the culture endpoint. Within the microfluidic chamber, an array of 3D tissue aggregates (spheroids) can be formed and cultured under perfusion. Subsequently, detailed post-culture analysis of the spheroids collected from the disassembled device can be performed. This device facilitates uniform spheroid formation, growth analysis in a high-throughput format, controlled proliferation via perfusion flow rate, and post-culture analysis of spheroids. We used the device to culture spheroids of human hepatocellular carcinoma (HepG2) cells under two controlled perfusion flow rates. HepG2 spheroids exhibited greater cell growth at higher perfusion flow rates than at lower perfusion flow rates, and exhibited different metabolic activity and mRNA and protein expression under the different flow rate conditions. These results show the potential of perfusion culture to precisely control the culture environment in microfluidic devices. The construction of spheroid array chambers allows multiple culture conditions to be tested simultaneously, with potential applications in toxicity and drug screening.