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Open AccessArticle

Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids

1
The Institute for Drug Research, The School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 91120, Israel
2
Department of Surgery and Transplantation Unit, Hadassah-Hebrew University Medical Center, Ein Kerem, Jerusalem 91120, Israel
3
Sharett Institute of Oncology, Hadassah-Hebrew University Medical Center, Ein Kerem, Jerusalem 91120, Israel
4
The Core Research Facility, The Hebrew University of Jerusalem, Jerusalem 91120, Israel
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2020, 21(20), 7703; https://doi.org/10.3390/ijms21207703
Received: 25 September 2020 / Revised: 11 October 2020 / Accepted: 16 October 2020 / Published: 18 October 2020
The currently accepted imaging methods have been a central hurdle to imaging the finer details of tumor behavior in three-dimensional (3D) ex vivo multicellular culture models. In our search for an improved way of imaging tumor behavior in its physiological-like niche, we developed a simple, efficient, and straightforward procedure using standard reagents and imaging equipment that significantly enhanced 3D imaging up to a ~200-micron depth. We tested its efficacy on pancreatic spheroids, prototypes of high-density tissues that are difficult to image. We found we could both save time with this method and extract information about pancreatic tumor spheroids that previously was difficult to obtain. We were able to discern clear differences in the organization of pancreatic tumor spheroids generated from different origins, suggesting cell-specific, inherent, bottom-up organization with a correlation to the level of malignancy. We also examined the dynamic changes in the spheroids at predetermined time points, providing important information related to tissue morphogenesis and its metabolic state. Lastly, this process enabled us to assess a drug vehicle’s potential to penetrate dense tumor tissue by improving our view of the inert particles’ diffusion in the 3D spheroid. This clearing method, a simple procedure, can open the door to more accurate imaging and reveal more about cancer behavior. View Full-Text
Keywords: spheroids; clearing; 3D imaging; pancreatic cancer spheroids; clearing; 3D imaging; pancreatic cancer
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Steinberg, E.; Orehov, N.; Tischenko, K.; Schwob, O.; Zamir, G.; Hubert, A.; Manevitch, Z.; Benny, O. Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids. Int. J. Mol. Sci. 2020, 21, 7703.

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