CoChat
Sign Up
Cited by 40Pluripotent Stem Cells ResearchDrug-Induced Hepatotoxicity and ProtectionMesenchymal stem cell research

Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study

Open access
Gao, Xiugong, Li, Rong, Cahan, Patrick, Zhao, Yang, Yourick, Jeffrey J., Sprando, Robert L.·2020·Stem Cell Research & Therapy
DOI 10.1186/s13287-020-01914-1PMC PMC7488531
Open this paper in CoChat — free accountOpen PDF

Abstract

Background: Hepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) hold great promise in toxicological applications as well as in regenerative medicine. Previous efforts on hepatocyte differentiation have mostly relied on the use of growth factors (GFs) to recapitulate developmental signals under in vitro conditions. Recently, the use of small molecules (SMs) has emerged as an attractive tool to induce cell fate transition due to its superiority in terms of both quality and cost. However, HLCs derived using SMs have not been well characterized, especially on the transcriptome level. Methods: HLCs were differentiated from human iPSCs using a protocol that only involves SMs and characterized by transcriptomic analysis using whole genome microarrays. Results: HLCs derived using the SM protocol (HLC_SM) displayed specific hepatic marker expression and demonstrated key hepatic functions. Transcriptomic analysis of the SM-driven differentiation defined a hepatocyte differentiation track and characterized the expression of some key marker genes in major stages of hepatocyte differentiation. In addition, HLC_SM were scored with CellNet, a bioinformatics tool quantifying how closely engineered cell populations resemble their target cell type, and compared to primary human hepatocytes (PHHs), adult liver tissue, fetal liver tissue, HLCs differentiated using GFs (HLC_GF), and commercially available HLCs. Similar to HLC_GF, HLC_SM displayed a mixed phenotype of fetal and adult hepatocytes and had relatively low expression of metabolic enzymes, transporters, and nuclear receptors compared to PHHs. Finally, the differentially expressed genes in HLC_SM compared to HLC_GF and to PHHs were analyzed to identify pathways and upstream transcription regulators which could potentially be manipulated to improve the differentiation of HLCs. Conclusions: Overall, the present study demonstrated the usefulness of the SM-based hepatocyte differentiation method, offered new insights into the molecular basis of hepatogenesis and associated gene regulation, and suggested ways for further improvements in hepatocyte differentiation in order to obtain more mature HLCs that could be used in toxicological studies.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1186/s13287-020-01914-1

Cites

5 of 5 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • The differentiation protocol used was developed and optimized from published literature and chosen for its capacity to generate iHLOs with high CYP enzyme activity, particularly CYP3A4.

    “The differentiation protocol utilized was developed and optimized from the published literatures18–21, specifically chosen for its capacity to generate iHLOs with high CYP enzyme activity, particularly CYP3A4, which is crucial for drug metabolism and toxicology evaluations.”

    From ​Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [21]

    The Methods section (‘Hepatocyte differentiation by only SMs’; page unavailable) says the authors followed a published protocol with slight modifications. The Results (‘Hepatic functions of HLCs derived by SMs’; page unavailable) report CYP3A activity in hepatocyte-like cells, but not iHLOs or specifically high CYP3A4 activity; the Results on drug-metabolizing enzymes report lower CYP3A4 expression than in primary hepatocytes and adult liver. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • The protocol was chosen due to its demonstrated capacity to generate iHLOs with high CYP enzyme activity, particularly CYP3A4.

    “This protocol was chosen due to its demonstrated capacity to generate iHLOs with high CYP enzyme activity, particularly CYP3A4, which is crucial for our subsequent compound or drug metabolism and toxicology evaluations.”

    From ​Temporal proteomic profiling of iPSC-derived human liver organoids reveals optimal maturation for drug metabolism and toxicology (2026) · cites this paper as [21]

    The Results section, “Hepatic functions of HLCs derived by SMs” (page unavailable), reports CYP3A activity in small-molecule-derived hepatocyte-like cells, comparable to growth-factor-derived cells. It does not report generating iHLOs or demonstrate high CYP3A4-specific activity. The Results section on drug-metabolizing enzymes instead reports lower CYP3A4 expression in these cells than in primary human hepatocytes and adult liver. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 32

  1. [1]Tolosa L Pareja E Gomez-Lechon MJ Clinical application of pluripotent stem cells: an alternative cell-based therapy for treating liver diseases? Transplantation. 2016 100 12 2548 2557 10.1097/TP.0000000000001426 27495745 · DOI
  2. [2]Davidson MD Ware BR Khetani SR Stem cell-derived liver cells for drug testing and disease modeling Discov Med 2015 19 106 349 358 26105698 PMC5768200
  3. [3]Goldring C Antoine DJ Bonner F Crozier J Denning C Fontana RJ Stem cell-derived models to improve mechanistic understanding and prediction of human drug-induced liver injury Hepatology (Baltimore, Md) 2017 65 2 710 721 10.1002/hep.28886 PMC5266558 27775817 · DOI
  4. [4]Takayama K Mizuguchi H Generation of human pluripotent stem cell-derived hepatocyte-like cells for drug toxicity screening Drug Metabolism Pharmacokinetics 2017 32 1 12 20 10.1016/j.dmpk.2016.10.408 28012798 · DOI
  5. [6]Krueger W Boelsterli UA Rasmussen TP Stem cell strategies to evaluate idiosyncratic drug-induced liver injury J Clin Transl Hepatol 2014 2 3 143 152 26355943 PMC4521249
  6. [7]Dornbos P LaPres JJ Incorporating population-level genetic variability within laboratory models in toxicology: from the individual to the population Toxicology. 2018 395 1 8 10.1016/j.tox.2017.12.007 29275117 PMC5801153 · DOI
  7. [8]Wetmore BA Allen B Clewell HJ 3rd Parker T Wambaugh JF Almond LM Incorporating population variability and susceptible subpopulations into dosimetry for high-throughput toxicity testing Toxicol Sci 2014 142 1 210 224 10.1093/toxsci/kfu169 25145659 · DOI
  8. [9]Schwartz RE Fleming HE Khetani SR Bhatia SN Pluripotent stem cell-derived hepatocyte-like cells Biotechnol Adv 2014 32 2 504 513 10.1016/j.biotechadv.2014.01.003 24440487 PMC4043206 · DOI
  9. [10]Palakkan AA Nanda J Ross JA Pluripotent stem cells to hepatocytes, the journey so far Biomed Reports 2017 6 4 367 373 10.3892/br.2017.867 PMC5374940 28413633 · DOI
  10. [11]Li X Xu J Deng H Small molecule-induced cellular fate reprogramming: promising road leading to Rome Curr Opin Genet Dev 2018 52 29 35 10.1016/j.gde.2018.05.004 29857280 · DOI
  11. [13]Gao X Yourick JJ Sprando RL Generation of nine induced pluripotent stem cell lines as an ethnic diversity panel Stem Cell Res 2018 31 193 196 10.1016/j.scr.2018.07.013 30099336 · DOI
  12. [14]Asplund A Pradip A van Giezen M Aspegren A Choukair H Rehnstrom M One standardized differentiation procedure robustly generates homogenous hepatocyte cultures displaying metabolic diversity from a large panel of human pluripotent stem cells Stem Cell Rev 2016 12 1 90 104 10.1007/s12015-015-9621-9 26385115 · DOI
  13. [15]Schneider CA Rasband WS Eliceiri KW NIH image to ImageJ: 25 years of image analysis Nat Methods 2012 9 7 671 675 10.1038/nmeth.2089 22930834 PMC5554542 · DOI · Paper page
  14. [16]Irizarry RA Hobbs B Collin F Beazer-Barclay YD Antonellis KJ Scherf U Exploration, normalization, and summaries of high density oligonucleotide array probe level data Biostatistics. 2003 4 2 249 264 10.1093/biostatistics/4.2.249 12925520 · DOI · Paper page
  15. [17]Tong W Cao X Harris S Sun H Fang H Fuscoe J ArrayTrack--supporting toxicogenomic research at the U.S. Food and Drug Administration National Center for Toxicological Research Environ Health Perspect 2003 111 15 1819 1826 10.1289/ehp.6497 14630514 PMC1241745 · DOI
  16. [18]Graffmann N Ncube A Wruck W Adjaye J Cell fate decisions of human iPSC-derived bipotential hepatoblasts depend on cell density PLoS One 2018 13 7 e0200416 10.1371/journal.pone.0200416 29990377 PMC6039024 · DOI
  17. [19]Cahan P, Li H, Morris SA, Lummertz da Rocha E, Daley GQ, Collins JJ. CellNet: network biology applied to stem cell engineering. Cell. 2014;158(4):903–915. PMC4233680 25126793
  18. [21]McCoy AT Benoist CC Wright JW Kawas LH Bule-Ghogare JM Zhu M Evaluation of metabolically stabilized angiotensin IV analogs as procognitive/antidementia agents J Pharmacol Exp Ther 2013 344 1 141 154 10.1124/jpet.112.199497 23055539 PMC3533412 · DOI
  19. [23]Gao X Liu Y A transcriptomic study suggesting human iPSC-derived hepatocytes potentially offer a better in vitro model of hepatotoxicity than most hepatoma cell lines Cell Biol Toxicol 2017 33 4 407 421 10.1007/s10565-017-9383-z 28144825 · DOI
  20. [24]Koturbash I Scherhag A Sorrentino J Sexton K Bodnar W Swenberg JA Epigenetic mechanisms of mouse interstrain variability in genotoxicity of the environmental toxicant 1,3-butadiene Toxicol Sci 2011 122 2 448 456 10.1093/toxsci/kfr133 21602187 PMC3155089 · DOI
  21. [25]Lu J Einhorn S Venkatarangan L Miller M Mann DA Watkins PB Morphological and functional characterization and assessment of iPSC-derived hepatocytes for in vitro toxicity testing Toxicol Sci 2015 147 1 39 54 10.1093/toxsci/kfv117 26092927 · DOI
  22. [26]Ward JH Jr Hierarchical grouping to optimize an objective function J Am Stat Assoc 1963 58 236 244 10.1080/01621459.1963.10500845 · DOI
  23. [27]Song Z Cai J Liu Y Zhao D Yong J Duo S Efficient generation of hepatocyte-like cells from human induced pluripotent stem cells Cell Res 2009 19 11 1233 1242 10.1038/cr.2009.107 19736565 · DOI · Paper page
  24. [28]Si-Tayeb K Noto FK Nagaoka M Li J Battle MA Duris C Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells Hepatology (Baltimore, Md) 2010 51 1 297 305 10.1002/hep.23354 PMC2946078 19998274 · DOI · Paper page
  25. [30]Touboul T Hannan NR Corbineau S Martinez A Martinet C Branchereau S Generation of functional hepatocytes from human embryonic stem cells under chemically defined conditions that recapitulate liver development Hepatology (Baltimore, Md) 2010 51 5 1754 1765 10.1002/hep.23506 20301097 · DOI
  26. [31]Raju R Chau D Notelaers T Myers CL Verfaillie CM Hu WS In vitro pluripotent stem cell differentiation to hepatocyte ceases further maturation at an equivalent stage of E15 in mouse embryonic liver development Stem Cells Dev 2018 27 13 910 921 10.1089/scd.2017.0270 29851366 PMC6916117 · DOI
  27. [32]Parviz F Matullo C Garrison WD Savatski L Adamson JW Ning G Hepatocyte nuclear factor 4alpha controls the development of a hepatic epithelium and liver morphogenesis Nat Genet 2003 34 3 292 296 10.1038/ng1175 12808453 · DOI · Paper page
  28. [33]Takayama K Inamura M Kawabata K Katayama K Higuchi M Tashiro K Efficient generation of functional hepatocytes from human embryonic stem cells and induced pluripotent stem cells by HNF4alpha transduction Molecular Therapy 2012 20 1 127 137 10.1038/mt.2011.234 22068426 PMC3255576 · DOI
  29. [34]Duncan SA Manova K Chen WS Hoodless P Weinstein DC Bachvarova RF Expression of transcription factor HNF-4 in the extraembryonic endoderm, gut, and nephrogenic tissue of the developing mouse embryo: HNF-4 is a marker for primary endoderm in the implanting blastocyst Proc Natl Acad Sci U S A 1994 91 16 7598 7602 10.1073/pnas.91.16.7598 8052626 PMC44449 · DOI
  30. [36]Godoy P Schmidt-Heck W Natarajan K Lucendo-Villarin B Szkolnicka D Asplund A Gene networks and transcription factor motifs defining the differentiation of stem cells into hepatocyte-like cells J Hepatol 2015 63 4 934 942 10.1016/j.jhep.2015.05.013 26022688 PMC4580233 · DOI
  31. [37]Baxter M Withey S Harrison S Segeritz CP Zhang F Atkinson-Dell R Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes J Hepatol 2015 62 3 581 589 10.1016/j.jhep.2014.10.016 25457200 PMC4334496 · DOI
  32. [38]Chen C Soto-Gutierrez A Baptista PM Spee B Biotechnology challenges to in vitro maturation of hepatic stem cells Gastroenterology. 2018 154 5 1258 1272 10.1053/j.gastro.2018.01.066 29428334 PMC6237283 · DOI

Cited by

Sorted by citations

Authors
Gao, X., Li, R., Cahan, P., Zhao, Y., Yourick, J. J., & Sprando, R. L.
Affiliations
  • Center for Food Safety and Applied Nutrition (US)
  • Johns Hopkins University (US)
Corresponding author
Gao, Xiugong
ORCID
Venue
Stem Cell Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2020-09-11
Volume
11
Issue
1
Pages
393-393
ISSN
1757-6512 (ISSN-L)
DOI
10.1186/s13287-020-01914-1
PMC
PMC7488531
OpenAlex
W3085363552
Cited by
40
References
5
Citations by year
2026: 2 · 2025: 14 · 2024: 5 · 2023: 8 · 2022: 8 · 2021: 3
FWCI
1.54
Type
journal article
Language
en
Open access
Licence
Locations
  • Stem Cell Research & Therapy (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Universitetet i Oslo
  • U.S. Food and Drug Administration
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
  • Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
Keywords
hepatocyte-like cells, induced pluripotent stem cells, small molecule differentiation, hepatocyte differentiation, transcriptome, hepatogenesis, primary human hepatocytes, metabolic enzymes, nuclear receptors, toxicology, regenerative medicine
MeSH terms
Adult, Cell Differentiation, Humans, Computational Biology, Hepatocytes, Induced Pluripotent Stem Cells, Transcriptome

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review