TY - JOUR
T1 - High-Throughput Screening Enhances Kidney Organoid Differentiation from Human Pluripotent Stem Cells and Enables Automated Multidimensional Phenotyping
AU - Czerniecki, Stefan M.
AU - Cruz, Nelly M.
AU - Harder, Jennifer L.
AU - Menon, Rajasree
AU - Annis, James
AU - Otto, Edgar A.
AU - Gulieva, Ramila E.
AU - Islas, Laura V.
AU - Kim, Yong Kyun
AU - Tran, Linh M.
AU - Martins, Timothy J.
AU - Pippin, Jeffrey W.
AU - Fu, Hongxia
AU - Kretzler, Matthias
AU - Shankland, Stuart J.
AU - Himmelfarb, Jonathan
AU - Moon, Randall T.
AU - Paragas, Neal
AU - Freedman, Benjamin S.
N1 - Publisher Copyright:
© 2018 Elsevier Inc.
PY - 2018/6/1
Y1 - 2018/6/1
N2 - Organoids derived from human pluripotent stem cells are a potentially powerful tool for high-throughput screening (HTS), but the complexity of organoid cultures poses a significant challenge for miniaturization and automation. Here, we present a fully automated, HTS-compatible platform for enhanced differentiation and phenotyping of human kidney organoids. The entire 21-day protocol, from plating to differentiation to analysis, can be performed automatically by liquid-handling robots, or alternatively by manual pipetting. High-content imaging analysis reveals both dose-dependent and threshold effects during organoid differentiation. Immunofluorescence and single-cell RNA sequencing identify previously undetected parietal, interstitial, and partially differentiated compartments within organoids and define conditions that greatly expand the vascular endothelium. Chemical modulation of toxicity and disease phenotypes can be quantified for safety and efficacy prediction. Screening in gene-edited organoids in this system reveals an unexpected role for myosin in polycystic kidney disease. Organoids in HTS formats thus establish an attractive platform for multidimensional phenotypic screening. Organoids derived from human iPSCs have great potential for drug screening, but their complexity poses a challenge for miniaturization and automation. Freedman and colleagues establish a robotic pipeline to manufacture and analyze kidney organoids in microwell arrays. They apply this system to improve differentiation, measure toxicity, and comprehend disease.
AB - Organoids derived from human pluripotent stem cells are a potentially powerful tool for high-throughput screening (HTS), but the complexity of organoid cultures poses a significant challenge for miniaturization and automation. Here, we present a fully automated, HTS-compatible platform for enhanced differentiation and phenotyping of human kidney organoids. The entire 21-day protocol, from plating to differentiation to analysis, can be performed automatically by liquid-handling robots, or alternatively by manual pipetting. High-content imaging analysis reveals both dose-dependent and threshold effects during organoid differentiation. Immunofluorescence and single-cell RNA sequencing identify previously undetected parietal, interstitial, and partially differentiated compartments within organoids and define conditions that greatly expand the vascular endothelium. Chemical modulation of toxicity and disease phenotypes can be quantified for safety and efficacy prediction. Screening in gene-edited organoids in this system reveals an unexpected role for myosin in polycystic kidney disease. Organoids in HTS formats thus establish an attractive platform for multidimensional phenotypic screening. Organoids derived from human iPSCs have great potential for drug screening, but their complexity poses a challenge for miniaturization and automation. Freedman and colleagues establish a robotic pipeline to manufacture and analyze kidney organoids in microwell arrays. They apply this system to improve differentiation, measure toxicity, and comprehend disease.
UR - http://www.scopus.com/inward/record.url?scp=85047213971&partnerID=8YFLogxK
U2 - 10.1016/j.stem.2018.04.022
DO - 10.1016/j.stem.2018.04.022
M3 - Article
C2 - 29779890
AN - SCOPUS:85047213971
SN - 1934-5909
VL - 22
SP - 929-940.e4
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 6
ER -