Hepatic differentiation of human iPSCs in different 3D models: A comparative study

Meier, Florian; Freyer, Nora; Brzeszczyńska, Joanna; Knöspel, Fanny; Armstrong, Lyle; Lako, Majlinda; Greuel, Selina; Damm, Georg; Ludwig-Schwellinger, Eva; Deschl, Ulrich; Ross, James A.; Beilmann, Mario; Zeilinger, Katrin

doi:10.3892/ijmm.2017.3190

Cited by 33 publications

(38 citation statements)

References 53 publications

(65 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, our experiments also call attention to the instability and as a consequence to the limited applicability of the commonly used liver model cell line, HepG2. Previous observations [38,56] and the data presented here suggest that the generally applied criteria for the differentiated HLCs are insufficient. Considering that polarized and functional expression of hepatic transporters is essential for liver functions [2,3,13,15,17,18], we recommend an augmentation of the usual tests with the assessment of the functional presence and proper localization of these transporter proteins when the stem cell-derived hepatic model systems are evaluated.…”

Section: Discussionmentioning

confidence: 55%

“…Drug inducibility of CYP enzymes is also an important indicator of drug metabolizing capacity of the hepatic cells; however, this feature is beyond a scope of our recent study. Previously, a few studies investigated the basal and drug-induced activities of CYPs in stem cell-derived HLCs, and demonstrated noticeable CYP activities that were further stimulated by drugs [38,56]. It should, however, be noted that these activities were 1000-fold smaller than found in primary hepatocytes.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

The importance of transporters and cell polarization for the evaluation of human stem cell-derived hepatic cells

et al. 2020

View full text Add to dashboard Cite

One of the most promising applications of human pluripotent stem cells is their utilization for human-based pharmacological models. Despite the fact that membrane transporters expressed in the liver play pivotal role in various hepatic functions, thus far only little attention was devoted to the membrane transporter composition of the stem cell-derived liver models. In the present work, we have differentiated HUES9, a human embryonic stem cell line, toward the hepatic lineage, and monitored the expression levels of numerous differentiation marker and liver transporter genes with special focus on ABC transporters. In addition, the effect of bile acid treatment and polarizing culturing conditions on hepatic maturation has been assessed. We found that most transporter genes crucial for hepatic functions are markedly induced during hepatic differentiation; however, as regards the transporter composition the end-stage cells still exhibited dual, hepatocyte and cholangiocyte character. Although the bile acid treatment and sandwich culturing only slightly influenced the gene expressions, the stimulated cell polarization resulted in formation of bile canaliculi and proper localization of transporters. Our results point to the importance of membrane transporters in human stem cell-derived hepatic models and demonstrate the relevance of cell polarization in generation of applicable cellular models with correctly localized transporters. On the basis of our observations we suggest that conventional criteria for the evaluation of the quality of stem cell-derived hepatocyte-like cells ought to be augmented with additional elements, such as polarized and functional expression of hepatic transporters.

show abstract

Section: Discussionmentioning

confidence: 55%

Section: Discussionmentioning

confidence: 99%

The importance of transporters and cell polarization for the evaluation of human stem cell-derived hepatic cells

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Even though nowadays it is possible to culture PHH for several days with preservation of their metabolic capacities in certain culture conditions, none of the above‐mentioned studies have shown that the appropriate measures have been taken to maintain PHH functions. The only three published studies that used fresh hepatocytes or hepatocytes cultured for maximally 24 hr as a control are the study of Ulvestad et al (), Freyer et al (), and Meier et al (). Comparison of the results of those studies with our data shows that the CYP3A, CYP2C9, and CYP1A activities of hiPSC‐derived hepatocytes in our study were 10 to several hundred folds higher.…”

Section: Discussionmentioning

confidence: 99%

Differentiation of human-induced pluripotent stem cell under flow conditions to mature hepatocytes for liver tissue engineering

Starokozhko

Hemmingsen

Larsen

et al. 2018

J Tissue Eng Regen Med

View full text Add to dashboard Cite

Hepatic differentiation of human‐induced pluripotent stem cells (hiPSCs) under flow conditions in a 3D scaffold is expected to be a major step forward for construction of bioartificial livers. The aims of this study were to induce hepatic differentiation of hiPSCs under perfusion conditions and to perform functional comparisons with fresh human precision‐cut liver slices (hPCLS), an excellent benchmark for the human liver in vivo. The majority of the mRNA expression of CYP isoenzymes and transporters and the tested CYP activities, Phase II metabolism, and albumin, urea, and bile acid synthesis in the hiPSC‐derived cells reached values that overlap those of hPCLS, which indicates a higher degree of hepatic differentiation than observed until now. Differentiation under flow compared with static conditions had a strong inducing effect on Phase II metabolism and suppressed AFP expression but resulted in slightly lower activity of some of the Phase I metabolism enzymes. Gene expression data indicate that hiPSCs differentiated into both hepatic and biliary directions. In conclusion, the hiPSC differentiated under flow conditions towards hepatocytes express a wide spectrum of liver functions at levels comparable with hPCLS indicating excellent future perspectives for the development of a bioartificial liver system for toxicity testing or as liver support device for patients.

show abstract

“…Differentiation of iPSCs involves cellular populations with compositions that progress along stages of the differentiation processes, [6][7][8] creating a naturally heterogeneous microenvironment with complex cellular interactions and, in 3D settings, these differentiating multicellular cultures have been demonstrated to develop spontaneously at the microscale. Although microengineered 3D cultures have been reported to improve differentiation quality, [9][10][11][12][13] owing to difficulties in handling 3D cultures, the field currently favors monolayer differentiation approaches. This approach may evolve with the development of microfabricated devices designed to perform with higher robustness and reliability for handling multicellular 3D microenvironments and for phenotyping their function in higher throughput settings.…”

Section: Impact Statementmentioning

confidence: 99%

Microengineered systems with iPSC-derived cardiac and hepatic cells to evaluate drug adverse effects

Dame

Ribeiro

2020

Exp Biol Med (Maywood)

View full text Add to dashboard Cite

Hepatic and cardiac drug adverse effects are among the leading causes of attrition in drug development programs, in part due to predictive failures of current animal or in vitro models. Hepatocytes and cardiomyocytes differentiated from human induced pluripotent stem cells (iPSCs) hold promise for predicting clinical drug effects, given their human-specific properties and their ability to harbor genetically determined characteristics that underlie inter-individual variations in drug response. Currently, the fetal-like properties and heterogeneity of hepatocytes and cardiomyocytes differentiated from iPSCs make them physiologically different from their counterparts isolated from primary tissues and limit their use for predicting clinical drug effects. To address this hurdle, there have been ongoing advances in differentiation and maturation protocols to improve the quality and use of iPSC-differentiated lineages. Among these are in vitro hepatic and cardiac cellular microsystems that can further enhance the physiology of cultured cells, can be used to better predict drug adverse effects, and investigate drug metabolism, pharmacokinetics, and pharmacodynamics to facilitate successful drug development. In this article, we discuss how cellular microsystems can establish microenvironments for these applications and propose how they could be used for potentially controlling the differentiation of hepatocytes or cardiomyocytes. The physiological relevance of cells is enhanced in cellular microsystems by simulating properties of tissue microenvironments, such as structural dimensionality, media flow, microfluidic control of media composition, and co-cultures with interacting cell types. Recent studies demonstrated that these properties also affect iPSC differentiations and we further elaborate on how they could control differentiation efficiency in microengineered devices. In summary, we describe recent advances in the field of cellular microsystems that can control the differentiation and maturation of hepatocytes and cardiomyocytes for drug evaluation. We also propose how future research with iPSCs within engineered microenvironments could enable their differentiation for scalable evaluations of drug effects. Impact statement Cardiac and hepatic adverse drug effects are among the leading causes of attrition in preclinical and clinical drug development programs as well as marketing withdrawals. The insufficiency of animal testing models has led to considerable interest in the employment of cardiac and hepatic models using human-induced pluripotent stem cells (iPSCs) for drug toxicity testing. However, current batches of iPSC-derived cardiomyocytes and hepatocytes are variable and not matured as adult primary tissues, which limit their prediction of drug effects. This article discusses how the use of microfluidics can create microenvironments to better control differentiation protocols and increase the physiological relevance of iPSC-derived cardiomyocytes and hepatocytes. Development and standardization of technologies will enable evaluation of the potential value of cellular microsystems to improve the in vitro models used in drug development programs. Future steps in this field include controlled connections of organ systems to better recreate clinical metabolism and pharmacokinetics.

show abstract

Hepatic differentiation of human iPSCs in different 3D models: A comparative study

Cited by 33 publications

References 53 publications

The importance of transporters and cell polarization for the evaluation of human stem cell-derived hepatic cells

The importance of transporters and cell polarization for the evaluation of human stem cell-derived hepatic cells

Differentiation of human-induced pluripotent stem cell under flow conditions to mature hepatocytes for liver tissue engineering

Microengineered systems with iPSC-derived cardiac and hepatic cells to evaluate drug adverse effects

Contact Info

Product

Resources

About