Ultra-High-Resolution 3D Optical Coherence Tomography Reveals Inner Structures of Human Placenta-Derived Trophoblast Organoids
Ultra-High-Resolution 3D Optical Coherence Tomography Reveals Inner Structures of Human Placenta-Derived Trophoblast Organoids
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DOI：
10.1109/tbme.2020.3038466
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发表时间：
2021-08-01
影响因子：
4.6
通讯作者：
Haindl, Richard
中科院分区：
文献类型：
--
作者：
Deloria, Abigail J.;Haider, Sandra;Haindl, Richard
目的：利用三维光学相干断层扫描（OCT）技术，在不进行样品制备的情况下，对人胎盘类器官进行原位分析。方法：分析的滋养层类器官来源于原代人滋养层。在这项研究中，一个定制的超高分辨率的光谱域OCT系统，具有均匀的空间和轴向分辨率为2.48 μ m的类器官组织中使用。获得的OCT结果与通过定量聚合酶链反应、蛋白质印迹分析、免疫组织化学和组织切片的免疫荧光检测的分化状态一致。结果：与明视野显微镜相比，3D OCT能够更详细地监测胎盘类器官。第一次在原位可视化和量化了腔体周围具有光散射“桥”的内部结构。这些桥和腔的形成与具有发育的合体滋养细胞的分化的滋养细胞类器官一致。结论：在活胎盘类器官中使用3D OCT是评估分化状态和原位解析内部结构的快速工具，这是标准活细胞成像模式无法实现的。重要性：直到最近才建立了人类胎盘来源的类器官，从而能够具有高度可重复和稳定的体外模型，不仅可以研究妊娠早期的发育过程，还可以研究妊娠早期的生理和病理生理过程。据我们所知，这项工作是第一次使用3D OCT分析活体人类胎盘类器官。因此，快速，特别是非终点OCT定性分析与类器官的分化阶段相一致，这将有助于该领域的未来发展。
Objective: 3D optical coherence tomography (OCT) is used for analyses of human placenta organoids in situ without sample preparation. Methods: The trophoblast organoids analyzed were derived from primary human trophoblast. In this study a custom made ultra-high-resolution spectral domain OCT system with uniform spatial and axial resolution of 2.48 mu m in organoid tissue was used. The obtained OCT results align to differentiation status tested via quantitative polymerase chain reaction, Western blot analyses, immunohistochemistry, and immunofluorescence of histological sections. Results: 3D OCT enables a more detailed placenta organoid monitoring compared to brightfield microscopy. Inner architecture with light scattering "bridges" surrounding cavities were visualized and quantified in situ for the first time. The formation of these bridges and cavities is congruent to differentiated trophoblast organoids having developed syncytiotrophoblasts. Conclusion: Using 3D OCT in living placenta organoids is a fast tool to assess the differentiation status and resolve internal structures in situ, which is not possible with standard live cell imaging modality. Significance: Only recently human placenta-derived organoids were established, allowing to have a highly reproducible and stable in vitro model to investigate not only developmental but also physiological and pathophysiological processes during early pregnancy. To our knowledge, this work is the first to analyze living human placenta organoids using 3D OCT. Thereby, the rapid and especially non-endpoint OCT qualitative analyses align to the differentiation stage of organoids, which will aid future advancement in this field.