Development of human pluripotent stem cell-derived hepatic organoids as an alternative model for drug safety assessment

Kim, Hye Min; Im, Ilkyun; Jeon, Jang Su; Kang, Eun‐Hye; Lee, Hyang‐Ae; Jo, Seongyea; Kim, Ji-Woo; Woo, Dong-Hun; Choi, Young Jae; Kim, Hyo Jin; Han, Ji-Seok; Lee, Byoung‐Seok; Kim, Jong-Hoon; Kim, Sang Kyum; Park, Han-Jin

doi:10.1016/j.biomaterials.2022.121575

Cited by 27 publications

(38 citation statements)

References 71 publications

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“… 191 The applicability of hepatic organoids derived from human iPSCs for drug toxicity assessment was demonstrated by comprehensive functional analysis of CYP450-mediated drug metabolism. 339 These cases demonstrate the versatility of organoids and their consistency with in vivo therapy, and these validation results promote the ubiquitous role of organoids in drug selection and prediction for clinical treatment.…”

Section: Digestive System Organoid Models and Precision And Personali...mentioning

confidence: 72%

Applications of human organoids in the personalized treatment for digestive diseases

Wang

Guo

Jin

et al. 2022

Sig Transduct Target Ther

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Digestive system diseases arise primarily through the interplay of genetic and environmental influences; there is an urgent need in elucidating the pathogenic mechanisms of these diseases and deploy personalized treatments. Traditional and long-established model systems rarely reproduce either tissue complexity or human physiology faithfully; these shortcomings underscore the need for better models. Organoids represent a promising research model, helping us gain a more profound understanding of the digestive organs; this model can also be used to provide patients with precise and individualized treatment and to build rapid in vitro test models for drug screening or gene/cell therapy, linking basic research with clinical treatment. Over the past few decades, the use of organoids has led to an advanced understanding of the composition of each digestive organ and has facilitated disease modeling, chemotherapy dose prediction, CRISPR-Cas9 genetic intervention, high-throughput drug screening, and identification of SARS-CoV-2 targets, pathogenic infection. However, the existing organoids of the digestive system mainly include the epithelial system. In order to reveal the pathogenic mechanism of digestive diseases, it is necessary to establish a completer and more physiological organoid model. Combining organoids and advanced techniques to test individualized treatments of different formulations is a promising approach that requires further exploration. This review highlights the advancements in the field of organoid technology from the perspectives of disease modeling and personalized therapy.

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Section: Digestive System Organoid Models and Precision And Personali...mentioning

confidence: 72%

Applications of human organoids in the personalized treatment for digestive diseases

Wang

Guo

Jin

et al. 2022

Sig Transduct Target Ther

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“…However, the use of matrigel is a crucial impairment for its translation, since matrigel is a poorly defined animal-derived matrix causing animal protein contamination and reproducibility (lot variation) issues. Researchers are now working on development of alternative biomaterial, mainly to replace matrigel with synthetic polymers ( Gjorevski et al, 2016 ; Garreta et al, 2021 ) or even with biomaterials derived from decellularized extracellular matrix ( Cho et al, 2021 ; Kim et al, 2022 ). The synthetic polymers have the advantage of tunability, while the decellularized extracellular matrix shows similarity with molecular cues of native tissues.…”

Section: Organoids Recapitulating Models Of Diseasesmentioning

confidence: 99%

“…Several models have been developed ( He et al, 2020 ), including co-culture of human hepatic progenitor and stellate cells for the simulation of a fibrotic condition ( Leite et al, 2016 ). For drug testing these organoids must show metabolic competence, mostly evaluated by CYP induction and albumin secretion ( Kim et al, 2022 ). Recently, a bioprinted model of human hepatocyte-like cells derived from organoids was tested under exposition of a known hepatotoxic compound, showing an expected decrease in cell viability ( Bouwmeester et al, 2021 ).…”

Section: Organoids As Platforms For Drug Developmentmentioning

confidence: 99%

3D organ-on-a-chip: The convergence of microphysiological systems and organoids

Baptista

Porrini

Kronemberger

et al. 2022

Front. Cell Dev. Biol.

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Medicine today faces the combined challenge of an increasing number of untreatable diseases and fewer drugs reaching the clinic. While pharmaceutical companies have increased the number of drugs in early development and entering phase I of clinical trials, fewer actually successfully pass phase III and launch into the market. In fact, only 1 out of every 9 drugs entering phase I will launch. In vitro preclinical tests are used to predict earlier and better the potential of new drugs and thus avoid expensive clinical trial phases. The most recent developments favor 3D cell culture and human stem cell biology. These 3D humanized models known as organoids better mimic the 3D tissue architecture and physiological cell behavior of healthy and disease models, but face critical issues in production such as small-scale batches, greater costs (when compared to monolayer cultures) and reproducibility. To become the gold standard and most relevant biological model for drug discovery and development, organoid technology needs to integrate biological culture processes with advanced microtechnologies, such as microphysiological systems based on microfluidics technology. Microphysiological systems, known as organ-on-a-chip, mimic physiological conditions better than conventional cell culture models since they can emulate perfusion, mechanical and other parameters crucial for tissue and organ physiology. In addition, they reduce labor cost and human error by supporting automated operation and reduce reagent use in miniaturized culture systems. There is thus a clear advantage in combining organoid culture with microsystems for drug development. The main objective of this review is to address the recent advances in organoids and microphysiological systems highlighting crucial technologies for reaching a synergistic strategy, including bioprinting.

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“…S3). To support the survival and maturation of ECs during the following period of mLO generation (16 days), we also optimized the DM previously used to produce liver organoids from liver tissue and iPSCs [ 37 – 39 ] by adding VEGF and bFGF [ 54 , 55 ]. mLOs were produced with or without HscLCs in ULA 96-well plates in DM for 5 days and further differentiated in ULA 24-well plates for 11 days using the same culture medium.…”

Section: Resultsmentioning

confidence: 99%

“…2 A. BG01-derived HEs, ECs, and HscLCs were dissociated into single-cell suspensions by treatment with TrypLE (12604-021, Gibco) for 3 min and collected using DMEM (12100-046, Gibco) containing 10% FBS (16000-044, Gibco). 1.5 × 10 4 HE, 3 × 10 4 ECs, and 5 × 10 3 HscLCs were seeded together in ultra-low attachment 96-well plates (ULA PrimeSurface® 96U, MS-9096UZ, SIMADZU) in cold multilineage liver organoid differentiation medium [ 37 – 40 ] [DM, advanced DMEM/F12 media supplemented with 1 × HEPES (15630-080, Gibco), 1 × GlutaMAX (35050-061, Gibco), 1 × penicillin/streptomycin (15140-122, Gibco), 0.1 mg/ml BSA (A7096, Sigma), 1 × B27 supplement minus vitamin A (12587-010, Gibco), 25 ng/ml BMP7 (120-03p, Peprotech), 100 ng/ml FGF19 (100-32, Peprotech), 25 ng/ml HGF (CYT-244, Prospec), 10 nM (Leu15)-Gastrin I (G9145, Sigma), 1.25 mM N-acetyl-L-cysteine (A9165, Sigma), 0.5 μM A83-01 (2939, Tocris Bioscience), 10 μM DAPT (ab120633, Abcam), 3 μM dexamethasone (D4902, Sigma), 0.566 mg/ml growth factor-reduced Matrigel (M354230, Corning), 100 ng/ml VEGF-165 (100-20, Peprotech), 50 ng/ml bFGF (100-18b, Peprotech), and 10 μM ROCK inhibitor Y27632 (1254, Tocris Bioscience)]. On days 1 and 3, cold DM was added to the organoid culture.…”

Section: Methodsmentioning

confidence: 99%

Generation of multilineage liver organoids with luminal vasculature and bile ducts from human pluripotent stem cells via modulation of Notch signaling

Kim

Yoo

et al. 2023

Stem Cell Res Ther

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Background The generation of liver organoids recapitulating parenchymal and non-parenchymal cell interplay is essential for the precise in vitro modeling of liver diseases. Although different types of multilineage liver organoids (mLOs) have been generated from human pluripotent stem cells (hPSCs), the assembly and concurrent differentiation of multiple cell types in individual mLOs remain a major challenge. Particularly, most studies focused on the vascularization of mLOs in host tissue after transplantation in vivo. However, relatively little information is available on the in vitro formation of luminal vasculature in mLOs themselves. Methods The mLOs with luminal blood vessels and bile ducts were generated by assembling hepatic endoderm, hepatic stellate cell-like cells (HscLCs), and endothelial cells derived entirely from hPSCs using 96-well ultra-low attachment plates. We analyzed the effect of HscLC incorporation and Notch signaling modulation on the formation of both bile ducts and vasculature in mLOs using immunofluorescence staining, qRT-PCR, ELISA, and live-perfusion imaging. The potential use of the mLOs in fibrosis modeling was evaluated by histological and gene expression analyses after treatment with pro-fibrotic cytokines. Results We found that hPSC-derived HscLCs are crucial for generating functional microvasculature in mLOs. HscLC incorporation and subsequent vascularization substantially reduced apoptotic cell death and promoted the survival and growth of mLOs with microvessels. In particular, precise modulation of Notch signaling during a specific time window in organoid differentiation was critical for generating both bile ducts and vasculature. Live-cell imaging, a series of confocal scans, and electron microscopy demonstrated that blood vessels were well distributed inside mLOs and had perfusable lumens in vitro. In addition, exposure of mLOs to pro-fibrotic cytokines induced early fibrosis-associated events, including upregulation of genes associated with fibrotic induction and endothelial cell activation (i.e., collagen I, α-SMA, and ICAM) together with destruction of tissue architecture and organoid shrinkage. Conclusion Our results demonstrate that mLOs can reproduce parenchymal and non-parenchymal cell interactions and suggest that their application can advance the precise modeling of liver diseases in vitro.

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Development of human pluripotent stem cell-derived hepatic organoids as an alternative model for drug safety assessment

Cited by 27 publications

References 71 publications

Applications of human organoids in the personalized treatment for digestive diseases

Applications of human organoids in the personalized treatment for digestive diseases

3D organ-on-a-chip: The convergence of microphysiological systems and organoids

Generation of multilineage liver organoids with luminal vasculature and bile ducts from human pluripotent stem cells via modulation of Notch signaling

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