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High Throughput Pancreatic Islet Microtissue 3D Characterization for Compound Screening
Author(s) -
Villani Thomas,
Johnson Michael,
Gardner Graeme
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.547.3
Subject(s) - confocal microscopy , in vivo , dapi , islet , confocal , cell culture , pancreatic islets , in vitro , microbiology and biotechnology , cell , chemistry , biomedical engineering , biophysics , biology , insulin , medicine , biochemistry , apoptosis , genetics , geometry , mathematics , endocrinology
In the last few years several companies have developed 3D in vitro cell culture models and have demonstrated that these 3D models more accurately depict in vivo characteristics compared to traditional 2D monolayer cell culture. Because of this improved in vivo relevancy, these models are being rapidly adopted in the drug discovery space as they can more accurately predict in vivo efficacy and toxicological characteristics. Of the different types of 3D cell culture models being used today, pancreatic islet microtissues have been shown to be a powerful tool for assessing the effectiveness of compounds on diabetes. However, current imaging approaches (e.g. confocal microscopy, wide‐field microscopy) are limited in their ability to completely characterize these 3D models as light attenuation limits imaging depth to approx. 1–3 cell layers. Therefore, current imaging techniques are not able to completely characterize the effect of compounds on beta cell proliferation. Through this work, pancreatic islet microtissues of approx. 250 μm diameter were exposed to several compounds that induce beta cell proliferation and the microtissues were then stained for EdU or Ki67, glucagon, insulin and DAPI. The labeled microtissues were then rendered transparent with a rapid and plate‐compatible tissue clearing technique that allows for the microtissues to be imaged in their entirety using high content confocal imaging. It was shown through this work that the addition of a tissue clearing technique resulted in a 3‐fold increase in the number of cells characterized in the microtissues compared to traditional wide‐field or confocal microscopy. Support or Funding Information This work was support by Visikol Inc. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .