Kim Suryong, Ko Jihoon, Lee Seung-Ryeol, Park Dohyun, Park Seunghyuk, Jeon Noo Li
Department of Mechanical Engineering, Seoul National University, Seoul, Republic of Korea.
Institute of Advanced Machines and Design, Seoul National University, Seoul, Republic of Korea.
Biotechnol Bioeng. 2021 Jul;118(7):2524-2535. doi: 10.1002/bit.27765. Epub 2021 Apr 8.
In vitro models are becoming more advanced to truly present physiological systems while enabling high-throughput screening and analysis. Organ-on-a-chip devices provide remarkable results through the reconstruction of a three-dimensional (3D) cellular microenvironment although they need to be further developed in terms of multiple liquid patterning principle, material properties, and scalability. Here we present a 3D anchor-based microfluidic injection-molded plastic array culture platform (Anchor-IMPACT) that enables selective, space-intensive patterning of hydrogels using anchor-island for high-throughput angiogenesis evaluation model. Anchor-IMPACT consists of a central channel and an anchor-island, integrating the array into an abbreviated 96-well plate format with a standard microscope slide size. The anchor-island enables selective 3D cell patterning without channel-to-channel contact or any hydrogel injection port using an anchor structure unlike conventional culture compartment. The hydrogel was patterned into defined regions by spontaneous capillary flow under hydrophilic conditions. We configured multiple cell patterning structures to investigate the angiogenic potency of colorectal cancer cells in Anchor-IMPACT and the morphological properties of the angiogenesis induced by the paracrine effect were evaluated. In addition, the efficacy of anticancer drugs against angiogenic sprouts was verified by following dose-dependent responses. Our results indicate that Anchor-IMPACT offers not only a model of high-throughput experimentation but also an advanced 3D cell culture platform and can significantly improve current in vitro models while providing the basis for developing predictive preclinical models for biopharmaceutical applications.
体外模型正变得越来越先进,以真正呈现生理系统,同时实现高通量筛选和分析。芯片器官装置通过重建三维(3D)细胞微环境取得了显著成果,尽管它们在多液体图案化原理、材料特性和可扩展性方面还需要进一步发展。在此,我们展示了一种基于3D锚定的微流控注塑塑料阵列培养平台(Anchor-IMPACT),该平台利用锚定岛实现水凝胶的选择性、空间密集型图案化,用于高通量血管生成评估模型。Anchor-IMPACT由一个中央通道和一个锚定岛组成,将阵列集成到具有标准显微镜载玻片尺寸的简化96孔板形式中。与传统培养隔室不同,锚定岛利用锚定结构实现选择性3D细胞图案化,无需通道间接触或任何水凝胶注射端口。在亲水条件下,水凝胶通过自发毛细流动被图案化到特定区域。我们构建了多种细胞图案化结构,以研究Anchor-IMPACT中结肠癌细胞的血管生成能力,并评估旁分泌效应诱导的血管生成的形态学特性。此外,通过跟踪剂量依赖性反应,验证了抗癌药物对血管生成芽的疗效。我们的结果表明,Anchor-IMPACT不仅提供了一个高通量实验模型,还提供了一个先进的3D细胞培养平台,能够显著改进当前的体外模型,同时为开发用于生物制药应用的预测性临床前模型提供基础。