Patra Bishnubrata, Peng Yu-Sheng, Peng Chien-Chung, Liao Wei-Hao, Chen Yu-An, Lin Keng-Hui, Tung Yi-Chung, Lee Chau-Hwang
Research Center for Applied Sciences , Academia Sinica, Taipei 11529, Taiwan.
Institute of Physics , Academia Sinica, Taipei 11529, Taiwan.
Biomicrofluidics. 2014 Sep 9;8(5):052109. doi: 10.1063/1.4895568. eCollection 2014 Sep.
We developed a microfluidic device to culture cellular spheroids of controlled sizes and suitable for live cell imaging by selective plane illumination microscopy (SPIM). We cocultured human umbilical vein endothelial cells (HUVECs) within the spheroids formed by hepatocellular carcinoma cells, and studied the distributions of the HUVECs over time. We observed that the migration of HUVECs depended on the size of spheroids. In the spheroids of ∼200 μm diameters, HUVECs migrated outwards to the edges within 48 h; while in the spheroids of ∼250 μm diameters, there was no outward migration of the HUVECs up to 72 h. In addition, we studied the effects of pro-angiogenic factors, namely, vascular endothelial growth factor (VEGF) and fibroblast growth factor (β-FGF), on the migration of HUVECs in the carcinoma cell spheroid. The outward migration of HUVECs in 200 μm spheroids was hindered by the treatment with VEGF and β-FGF. Moreover, some of the HUVECs formed hollow lumen within 72 h under VEGF and β-FGF treatment. The combination of SPIM and microfluidic devices gives high resolution in both spatial and temporal domains. The observation of HUVECs in spheroids provides us insight on tumor vascularization, an ideal disease model for drug screening and fundamental studies.
我们开发了一种微流控装置，用于培养大小可控且适合通过选择性平面照明显微镜（SPIM）进行活细胞成像的细胞球体。我们将人脐静脉内皮细胞（HUVECs）与肝癌细胞形成的球体共培养，并研究了HUVECs随时间的分布情况。我们观察到HUVECs的迁移取决于球体的大小。在直径约为200μm的球体中，HUVECs在48小时内向外迁移至边缘；而在直径约为250μm的球体中，直到72小时HUVECs都没有向外迁移。此外，我们研究了促血管生成因子，即血管内皮生长因子（VEGF）和成纤维细胞生长因子（β-FGF）对癌细胞球体中HUVECs迁移的影响。用VEGF和β-FGF处理会阻碍200μm球体中HUVECs的向外迁移。此外，在VEGF和β-FGF处理下，一些HUVECs在72小时内形成了中空管腔。SPIM和微流控装置的结合在空间和时间域上都提供了高分辨率。对球体中HUVECs的观察为我们提供了关于肿瘤血管生成的见解，这是一种用于药物筛选和基础研究的理想疾病模型。
