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Automated System Developed for Human Organoid Production

By BiotechDaily International staff writers
Posted on 30 May 2018
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Image: Micrograph of a microwell plate containing kidney organoids, generated by liquid handling robots from human stem cells. Red, green, and yellow colors mark distinct segments of the kidney (Photo courtesy of the Freedman Laboratory, University of Washington School of Medicine).
Image: Micrograph of a microwell plate containing kidney organoids, generated by liquid handling robots from human stem cells. Red, green, and yellow colors mark distinct segments of the kidney (Photo courtesy of the Freedman Laboratory, University of Washington School of Medicine).
An automated liquid handling system has been established for the rapid production of human organoids derived from pluripotent stem cells.

Organoids derived from human pluripotent stem cells are a potentially powerful tool for use in cellular research utilizing high-throughput screening (HTS), but the complexity of maintaining organoid cultures has posed a significant challenge for miniaturization and automation.

In order to simplify working with organoids, investigators at the University of Washington School of Medicine (Seattle, USA) developed a fully automated, HTS-compatible platform for enhanced differentiation and phenotyping of human kidney organoids. This system relied on liquid-handling robots to seed pluripotent stem cells onto 384-well microtiter plates. Each microwell eventually generated ten or more organoids. The entire 21-day protocol, from plating to differentiation to analysis, was performed automatically by liquid-handling robots.

The investigators reported in the May 17, 2018, online edition of the journal Cell Stem Cell that high-content imaging analysis revealed both dose-dependent and threshold effects during organoid differentiation. Immunofluorescence and single-cell RNA sequencing identified previously undetected parietal, interstitial, and partially differentiated compartments within organoids and defined conditions that greatly expanded the vascular endothelium.

In an extension of the protocol, the investigators produced genetically engineered organoids carrying mutations that caused polycystic kidney disease, a common, inherited condition that affects one in 600 people worldwide and often leads to kidney failure. Screening these gene-edited organoids in this system revealed an unexpected role for myosin in polycystic kidney disease.

"This is a new "secret weapon" in our fight against disease," said senior author Dr. Benjamin Freedman, assistant professor of medicine at the University of Washington School of Medicine. "Ordinarily, just setting up an experiment of this magnitude would take a researcher all day, while the robot can do it in 20 minutes. On top of that, the robot does not get tired and make mistakes. "There is no question. For repetitive, tedious tasks like this, robots do a better job than humans."

"These findings give us a better idea of the nature of these organoids and provide a baseline from which we can make improvements," said Dr. Freedman. "The value of this high-throughput platform is that we can now alter our procedure at any point, in many different ways, and quickly see which of these changes produces a better result."

Related Links:
University of Washington School of Medicine


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