4D imaging and analysis of multicellular tumour spheroid cell migration and invasion

4D imaging and analysis of multicellular tumour spheroid cell migration and invasion
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多细胞肿瘤球状细胞迁移和侵袭的4D成像和分析

DOI:
10.1101/443648
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发表时间:
2018
期刊:
--
影响因子:
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通讯作者:
Richards R
Richards R
中科院分区:
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文献类型:
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作者:
Richards R

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研究和表征肿瘤细胞迁移对于更好地了解疾病进展和评估药物疗效至关重要。虽然肿瘤细胞迁移基本上发生在3个空间维度(3D),但由于生长3D细胞培养物的实用性以及随着时间的推移测量和跟踪3D运动的复杂性,迄今为止大多数迁移研究都在2D中进行分析。在这里,我们用光片荧光显微镜成像活的多细胞肿瘤球体,以确定细胞随时间的3D迁移和侵袭(4D)。我们专注于胶质母细胞瘤,这是一种侵袭性脑肿瘤,其中细胞侵入周围正常大脑仍然是临床管理的主要挑战。我们开发了一个分析复杂的3D细胞运动的工作流程,考虑到球体内的迁移以及侵入周围基质。这提供了总结不同实验条件下细胞运动的指标。作为原理的证明,我们展示了如何使用3D跟踪和分析来评估化疗药物对侵袭的疗效。丰富的数据集为进一步研究药物疗效,微环境组成和刚度以及集体细胞迁移和转移潜力开辟了途径。
Studying and characterising tumour cell migration is critical for a better understanding of disease progression and for assessing drug efficacy. Whilst tumour cell migration occurs fundamentally in 3 spatial dimensions (3D), because of the practicalities of growing 3D cell cultures and the complexities in measuring and tracking movement in 3D over time, most migration studies to date have performed analysis in 2D. Here we imaged live multicellular tumour spheroids with lightsheet fluorescence microscopy to determine cellular migration and invasion in 3D over time (4D). We focused on glioblastoma, which are aggressive brain tumours, where cell invasion into the surrounding normal brain remains a major challenge for clinical management. We developed a workflow for analysing complex 3D cell movement, taking into account migration within the spheroid as well as invasion into the surrounding matrix. This provided metrics that summarize the motion of the cells in different experimental conditions. As a proof of principle, we show how 3D tracking and analysis can be used to evaluate the efficacy of chemotherapeutics on invasion. The rich datasets open avenues for further studies on drug efficacy, microenvironment composition and stiffness, as well as collective cell migration and metastatic potential.
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