Beyond the surface: Investigation of tumorsphere morphology using volume electron microscopy

IF 3 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Nickhil Jadav , Sailakshmi Velamoor , Daniel Huang , Léna Cassin , Niki Hazelton , Alice-Roza Eruera , Laura N. Burga , Mihnea Bostina
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引用次数: 0

Abstract

The advent of volume electron microscopy (vEM) has provided unprecedented insights into cellular and subcellular organization, revolutionizing our understanding of cancer biology. This study presents a previously unexplored comparative analysis of the ultrastructural disparities between cancer cells cultured as monolayers and tumorspheres. By integrating a robust workflow that incorporates high-pressure freezing followed by freeze substitution (HPF/FS), serial block face scanning electron microscopy (SBF-SEM), manual and deep learning-based segmentation, and statistical analysis, we have successfully generated three-dimensional (3D) reconstructions of monolayer and tumorsphere cells, including their subcellular organelles. Our findings reveal a significant degree of variation in cellular morphology in tumorspheres. We observed the increased prevalence of nuclear envelope invaginations in tumorsphere cells compared to monolayers. Furthermore, we detected a diverse range of mitochondrial morphologies exclusively in tumorsphere cells, as well as intricate cellular interconnectivity within the tumorsphere architecture. These remarkable ultrastructural differences emphasize the use of tumorspheres as a superior model for cancer research due to their relevance to in vivo conditions. Our results strongly advocate for the utilization of tumorsphere cells in cancer research studies, enhancing the precision and relevance of experimental outcomes, and ultimately accelerating therapeutic advancements.

超越表面:使用体积电子显微镜研究肿瘤球体形态。
体积电子显微镜(vEM)的出现为细胞和亚细胞组织提供了前所未有的见解,彻底改变了我们对癌症生物学的理解。本研究对单层培养的癌症细胞和肿瘤球之间的超微结构差异进行了先前未经探索的比较分析。通过集成强大的工作流程,包括高压冷冻和冷冻替代(HPF/FS)、连续块面扫描电子显微镜(SBF-SEM)、手动和基于深度学习的分割以及统计分析,我们成功地生成了单层和肿瘤细胞层细胞的三维(3D)重建,包括其亚细胞器。我们的发现揭示了肿瘤球细胞形态的显著变化。我们观察到,与单层相比,肿瘤球细胞中核膜内陷的发生率增加。此外,我们在肿瘤层细胞中检测到了各种各样的线粒体形态,以及肿瘤层结构中复杂的细胞互连性。这些显著的超微结构差异强调了肿瘤球作为癌症研究的优越模型的使用,因为它们与体内条件相关。我们的研究结果强烈主张在癌症研究中利用肿瘤球细胞,提高实验结果的准确性和相关性,并最终加速治疗进展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of structural biology
Journal of structural biology 生物-生化与分子生物学
CiteScore
6.30
自引率
3.30%
发文量
88
审稿时长
65 days
期刊介绍: Journal of Structural Biology (JSB) has an open access mirror journal, the Journal of Structural Biology: X (JSBX), sharing the same aims and scope, editorial team, submission system and rigorous peer review. Since both journals share the same editorial system, you may submit your manuscript via either journal homepage. You will be prompted during submission (and revision) to choose in which to publish your article. The editors and reviewers are not aware of the choice you made until the article has been published online. JSB and JSBX publish papers dealing with the structural analysis of living material at every level of organization by all methods that lead to an understanding of biological function in terms of molecular and supermolecular structure. Techniques covered include: • Light microscopy including confocal microscopy • All types of electron microscopy • X-ray diffraction • Nuclear magnetic resonance • Scanning force microscopy, scanning probe microscopy, and tunneling microscopy • Digital image processing • Computational insights into structure
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