Leveraging interacting signaling pathways to robustly improve the quality and yield of human pluripotent stem cell-derived hepatoblasts and hepatocytes

Raggi, Claudia; M'Callum, Marie-Agnès; Pham, Quang Toan; Gaub, Perrine; Selleri, Silvia; Baratang, Nissan Vida; Mangahas, C.; Cagnone, Gaël; Reversade, Bruno; Joyal, Jean‐Sébastien; Paganelli, Massimiliano

doi:10.1016/j.stemcr.2022.01.003

Cited by 13 publications

(11 citation statements)

References 59 publications

(98 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Liver organoids are generated by seeding iPSC-derived hepatoblasts, endothelial progenitors, and mesenchymal progenitors on Matrigel in a fixed ratio. The organoids selfaggregate over 72 hr, reaching 1-2 mm in diameter, and show mature liver functions (Cyp3A4 activity, albumin secretion, urea production) 5 days after seeding (Raggi et al, 2022).…”

Section: Generation Of Complex Syngeneic Liver Organoidsmentioning

confidence: 99%

“…Protocols were progressively refined to generate more mature hepatocyte‐like cells (HLCs) from either embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) by acting on very diverse signaling pathways (Hannan, Segeritz, Touboul, & Vallier, 2013; McLean et al., 2007; Shan et al., 2013; Takayama et al., 2017; Touboul et al., 2016). We recently described a new robust differentiation protocol that allows for consistent generation of high‐quality, homogeneous, and functional hepatoblasts and HLCs in high yield from both human ESCs and iPSCs (Raggi et al., 2022). However, despite the heterogeneity of growth factors, small molecules, culture media, and coating compounds used, the full maturity of adult hepatocytes was never achieved in vitro with the described protocols, and a significant variability between various iPSC lines and between ESCs and iPSCs was often observed (Baxter et al., 2015; Kajiwara et al., 2012).…”

Section: Introductionmentioning

confidence: 99%

“…The shortcomings of such bidimensional models have pushed for the development of three‐dimensional (3D) culture systems that support the survival and maturation of liver cells (Hu et al., 2018). By recreating the interplay between hepatocytes and other cell types composing the liver microenvironment, organoids including mesenchymal and endothelial cells were shown to better recreate the cell‐cell interactions crucial for cell maturation (Asai et al., 2017; Berger, Ware, Davidson, Allsup, & Khetani, 2015; Raggi et al., 2022; Shinozawa, Yoshikawa, & Takebe, 2016; Takebe et al., 2013). We and others have showed that such 3D models better recapitulate the complexity of the tissue and enable, always within controlled experimental conditions, representative study of phenomena that guide human liver development or lead to complex liver diseases (Camp et al., 2017; Koike et al., 2019; Nie et al., 2018; Raggi et al., 2022).…”

Section: Introductionmentioning

confidence: 99%

“…Here we describe a robust protocol to generate complex human liver organoids composed of several cell types derived from a single iPSC population. Such organoids allow for the recreation of interactions between hepatocytes and endothelial and mesenchymal cells within the developing liver or in response to external stimuli (Raggi et al., 2022). Furthermore, being derived from a single iPSC population, all cell types share the same genetic background, which allows for the study of pathogenic consequences of patient‐specific genomes or, when coupled with genome editing of iPSCs, assessment of the effect of specific mutations on the development of complex liver diseases (Pham et al., 2020).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Generation of Complex Syngeneic Liver Organoids from Induced Pluripotent Stem Cells to Model Human Liver Pathophysiology

et al. 2022

Self Cite

View full text Add to dashboard Cite

The study of human liver pathophysiology has been hampered for decades by the lack of easily accessible, robust, and representative in vitro models. The discovery of induced pluripotent stem cells (iPSCs)—which can be generated from patients’ somatic cells, engineered to harbor specific mutations, and differentiated into hepatocyte‐like cells—opened the way to more meaningful modeling of liver development and disease. Nevertheless, representative modeling of many complex liver conditions requires the recreation of the interplay between hepatocytes and nonparenchymal liver cells. Here we describe protocols we developed to generate and characterize complex human liver organoids composed of iPSC‐derived hepatic, endothelial, and mesenchymal cells. With all cell types derived from the same iPSC population, such organoids reproduce the liver niche, allowing for the study of liver development and the modeling of complex inflammatory and fibrotic conditions. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Differentiation of human iPSCs into hepatic progenitor cells (hepatoblasts) Basic Protocol 2: Differentiation of human iPSCs into endothelial progenitor cells Support Protocol 1: Characterization of iPSC‐derived endothelial progenitor cells Basic Protocol 3: Differentiation of human iPSCs into mesenchymal progenitor cells Support Protocol 2: Characterization of iPSC‐derived mesenchymal progenitor cells Basic Protocol 4: Generation of complex syngeneic liver organoids

show abstract

Section: Generation Of Complex Syngeneic Liver Organoidsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Generation of Complex Syngeneic Liver Organoids from Induced Pluripotent Stem Cells to Model Human Liver Pathophysiology

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Similar to differentiation protocols for embryonic stem cells, hepatocyte differentiation from iPSCs is mimicked using a multistage cascade via endoderm, anterior definitive endoderm, and hepatocyte commitment through to hepatocyte-like cells (HLCs) ( Si-Tayeb et al, 2010 ; Touboul et al, 2010 ; Hannan et al, 2013 ; Mathapati et al, 2016 ). Failure to express drug metabolizing enzymes, at levels comparable to PHH ( Baxter et al, 2015 ; Sampaziotis et al, 2015 ) is being improved as the complex mimicry of liver development is better recapitulated by differentiation protocols which direct cellular fate with greater fidelity ( Ouchi et al, 2019 ; Raggi et al, 2022 ; Takeishi et al, 2020 ). In addition to these more physiologically relevant models, the improved benchmarking of liver metabolizing enzymes compared to both fetal and adult counterparts ( Zabulica et al, 2019 ), is better defining how HLCs can be appropriately applied to biological questions.…”

Section: Hepatocyte Models In Precision Medicinementioning

confidence: 99%

Hepatic Models in Precision Medicine: An African Perspective on Pharmacovigilance

2022

View full text Add to dashboard Cite

Pharmaceuticals are indispensable to healthcare as the burgeoning global population is challenged by diseases. The African continent harbors unparalleled genetic diversity, yet remains largely underrepresented in pharmaceutical research and development, which has serious implications for pharmaceuticals approved for use within the African population. Adverse drug reactions (ADRs) are often underpinned by unique variations in genes encoding the enzymes responsible for their uptake, metabolism, and clearance. As an example, individuals of African descent (14–34%) harbor an exclusive genetic variant in the gene encoding a liver metabolizing enzyme (CYP2D6) which reduces the efficacy of the breast cancer chemotherapeutic Tamoxifen. However, CYP2D6 genotyping is not required prior to dispensing Tamoxifen in sub-Saharan Africa. Pharmacogenomics is fundamental to precision medicine and the absence of its implementation suggests that Africa has, to date, been largely excluded from the global narrative around stratified healthcare. Models which could address this need, include primary human hepatocytes, immortalized hepatic cell lines, and induced pluripotent stem cell (iPSC) derived hepatocyte-like cells. Of these, iPSCs, are promising as a functional in vitro model for the empirical evaluation of drug metabolism. The scale with which pharmaceutically relevant African genetic variants can be stratified, the expediency with which these platforms can be established, and their subsequent sustainability suggest that they will have an important role to play in the democratization of stratified healthcare in Africa. Here we discuss the requirement for African hepatic models, and their implications for the future of pharmacovigilance on the African continent.

show abstract

Developing Liver Microphysiological Systems for Biomedical Applications

Wang,

Wu,

Zhao

et al. 2023

Adv Healthcare Materials

View full text Add to dashboard Cite

Microphysiological systems (MPSs), also known as organ chips, are micro‐units that integrate cells with diverse physical and biochemical environmental cues. In the field of liver MPSs, cellular components have advanced from simple planar cell cultures to more sophisticated 3D formations such as spheroids and organoids. Additionally, progress in microfluidic devices, bioprinting, engineering of matrix materials, and interdisciplinary technologies have significant promise for producing MPSs with biomimetic structures and functions. This review provides a comprehensive summary of biomimetic liver MPSs including their clinical applications and future developmental potential. First, the key components of liver MPSs, including the principal cell types and engineered structures utilized for cell cultivation, are briefly introduced. Subsequently, the biomedical applications of liver MPSs, including the creation of disease models, drug absorption, distribution, metabolism, excretion, and toxicity, are discussed. Finally, the challenges encountered by MPSs are summarized, and future research directions for their development are proposed.

show abstract

Leveraging interacting signaling pathways to robustly improve the quality and yield of human pluripotent stem cell-derived hepatoblasts and hepatocytes

Cited by 13 publications

References 59 publications

Generation of Complex Syngeneic Liver Organoids from Induced Pluripotent Stem Cells to Model Human Liver Pathophysiology

Generation of Complex Syngeneic Liver Organoids from Induced Pluripotent Stem Cells to Model Human Liver Pathophysiology

Hepatic Models in Precision Medicine: An African Perspective on Pharmacovigilance

Developing Liver Microphysiological Systems for Biomedical Applications

Contact Info

Product

Resources

About