Bioengineered Scaffolds for 3D Analysis of Glioblastoma Proliferation and Invasion
Bioengineered Scaffolds for 3D Analysis of Glioblastoma Proliferation and Invasion
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DOI：
10.1007/s10439-014-1223-1
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发表时间：
2015-08-01
影响因子：
3.8
通讯作者：
Sirianni, Rachael W.
中科院分区：
文献类型：
--
作者：
Heffernan, John M.;Overstreet, Derek J.;Sirianni, Rachael W.
恶性胶质母细胞瘤（GBM）细胞侵入健康脑是肿瘤复发和相关发病的主要原因。在这里，我们描述了一种高通量的方法，用于定量测量GBM增殖和侵袭的三维（3D）培养。由巯基化透明质酸和明胶组成的光学透明水凝胶与巯基反应性聚（乙二醇）聚合物化学交联以形成人工3D肿瘤微环境。黏弹性和水稳定性的表征表明，水凝胶具有机械可调性，硬度范围为18 Pa至18.2 kPa，并且抗水解至少30天。在水凝胶上培养的U118和U87R GBM球体的增殖、播散和随后的侵袭用重复的荧光共聚焦显微镜原位跟踪。使用定制的自动化图像处理，在14天内通过500 μ m凝胶鉴定和定量细胞。观察到细胞类型、凝胶硬度和肝细胞生长因子的可用性决定了细胞的增殖和侵袭行为。这些测量突出了该平台在人工3D肿瘤微环境中对恶性细胞行为进行定量荧光成像分析的实用性。
The invasion of malignant glioblastoma (GBM) cells into healthy brain is a primary cause of tumor recurrence and associated morbidity. Here, we describe a high-throughput method for quantitative measurement of GBM proliferation and invasion in three-dimensional (3D) culture. Optically clear hydrogels composed of thiolated hyaluronic acid and gelatin were chemically crosslinked with thiol-reactive poly(ethylene glycol) polymers to form an artificial 3D tumor microenvironment. Characterization of the viscoelasticity and aqueous stability indicated the hydrogels were mechanically tunable with stiffness ranging from 18 Pa to 18.2 kPa and were resistant to hydrolysis for at least 30 days. The proliferation, dissemination and subsequent invasion of U118 and U87R GBM spheroids cultured on the hydrogels were tracked in situ with repeated fluorescence confocal microscopy. Using custom automated image processing, cells were identified and quantified through 500 A mu m of gel over 14 days. Proliferative and invasive behaviors were observed to be contingent on cell type, gel stiffness, and hepatocyte growth factor availability. These measurements highlight the utility of this platform for performing quantitative, fluorescence imaging analysis of the behavior of malignant cells within an artificial, 3D tumor microenvironment.