Ultra-High-Resolution 3D Optical Coherence Tomography Reveals Inner Structures of Human Placenta-Derived Trophoblast Organoids.

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 μm 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.