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Human pluripotent stem cell-derived hepatic organoids as a
screening platform for toxicity prediction and drug evaluation

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Seon Ju Mun , Jaeseo Lee , Mi-Ok Lee , Ye Seul Son , Soo Jin Oh , Hyun-Soo Cho , Mi-Young Son , Dae-Soo Kim ,
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Ho-Joon Lee , Janghwan Kim , Cho-Rok Jung , Kyung-Sook Chung 1,2,5,* and Myung Jin Son 1,2, *
BACKGROUND AIM
Human cell-based and personalized in vitro liver models The development of hepatic models capable of long-
are urgently needed for drug efficacy and toxicity tests in term expansion with competent liver functionality in a
pre-clinical drug development. Although the liver is a personalized setting is technically challenging. Stem
representative organ with a native regenerative potential cell-based organoid technologies can provide an
in vivo, primary human hepatocytes (PHHs), which are alternative source of patient-derived primary
considered the gold standard for evaluating hepatic hepatocytes. However, self-renewing and functionally
metabolism, are limited by their loss of proliferative competent human pluripotent stem cell (PSC)-derived
capacity and long-term functionality in vitro. hepatic organoids are still lacking
METHODS
We developed a novel method to efficiently and reproducibly generate functionally mature human hepatic organoids
derived from PSCs, including human embryonic stem cells and induced PSCs. The maturity of the organoids was
validated by a detailed transcriptome analysis and functional performance assays. The organoids were applied to
screening platforms for predicting toxicity and evaluating drugs that target hepatic steatosis through real-time
monitoring of cellular bioenergetics and high-content analyses.
RESULTS

Our organoids were morphologically Figure 1 Figure 2
indistinguishable from adult liver tissue-derived
epithelial organoids and exhibited self-renewal.
With further maturation, their molecular
features approximated those of liver tissue,
although these features were lacking in two-
dimensionally differentiated hepatocytes. Our
organoids preserved mature liver properties,
including serum proteins production, drug
metabolism and detoxifying functions, active
mitochondrial bioenergetics, and regenerative
and inflammatory responses. The organoids
exhibited significant toxic responses to clinically
relevant concentrations of drugs that had been
withdrawn from the market due to hepa- Figure 3 Figure 4
totoxicity and recapitulated human disease
phenotypes such as hepatic steatosis.
Fig. 1. Generation and differentiation of
expandable human hepatic organoids from
PSCs.
Fig. 2. Transcriptome profiles and functional
assessment of human hepatic organoids
Fig. 3. Toxicological outcome prediction using
human hepatic organoids
Fig. 4. Human hepatic organoids for modeling
steatosis pathology and drug screening
CONCLUSION REFERENCES ACKNOWLEDGEMENTS
Our organoids exhibit self-renewal
(expandable and further able to • Takebe T, et al. Vascularized and This work was supported by the
differentiate) while maintaining their functional human liver from an KRIBB Initiative of the Korea
mature hepatic characteristics over iPSC-derived organ bud transplant. Research Council of Fundamental
long-term culture, and they may Nature 2013;499:481-484. Science and Technology
provide a versatile and valuable • Huch M, et al. Long-term culture of
platform for physiologically and genome-stable bipotent stem cells Contact information
pathologically relevant hepatic mo- from adult human liver. Cell Corresponding Author; Myung Jin Son
dels in the context of personalized 2015;160:299-312
medicine mjson@kribb.re.kr