Department of Life and Environment Engineering, The University of Kitakyushu, Kitakyushu, Fukuoka, Japan; Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
Biotechnol J. 2014 Jul;9(7):971-9. doi: 10.1002/biot.201300559. Epub 2014 Jun 12.
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.
微流控装置允许对三维（3D）组织进行灌注培养，模拟我们体内血管化 3D 组织中血液的流动。在这里，我们报告了一种由两部分微流控腔芯片和多微孔阵列芯片组成的微流控装置，该装置能够在培养终点处拆卸。在微流控腔室内，可以形成并培养 3D 组织聚集体（球体）的阵列，进行灌注培养。然后，可以对从拆开的装置中收集的球体进行详细的培养后分析。该装置有利于均匀地形成球体，以高通量格式进行生长分析，通过灌注流速来控制增殖，并对球体进行培养后分析。我们使用该装置在两种控制的灌注流速下培养人肝癌（HepG2）细胞的球体。与较低的灌注流速相比，HepG2 球体在较高的灌注流速下表现出更高的细胞生长速度，并且在不同的流速条件下表现出不同的代谢活性以及 mRNA 和蛋白质表达。这些结果表明，灌注培养具有精确控制微流控装置中培养环境的潜力。球体阵列腔室的构建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。
