Small-Molecule-Driven Hepatocyte Differentiation of Human Pluripotent Stem Cells
Open accessAbstract
The differentiation of pluripotent stem cells to hepatocytes is well established, yet current methods suffer from several drawbacks. These include a lack of definition and reproducibility, which in part stems from continued reliance on recombinant growth factors. This has remained a stumbling block for the translation of the technology into industry and the clinic for reasons associated with cost and quality. We have devised a growth-factor-free protocol that relies on small molecules to differentiate human pluripotent stem cells toward a hepatic phenotype. The procedure can efficiently direct both human embryonic stem cells and induced pluripotent stem cells to hepatocyte-like cells. The final population of cells demonstrates marker expression at the transcriptional and protein levels, as well as key hepatic functions such as serum protein production, glycogen storage, and cytochrome P450 activity.
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Growth factors can be costly and unstable, and many small-molecule drugs affect multiple pathways, resulting in poor reproducibility.
“While growth factors could be costly and unstable, and many small-molecule drugs can affect multiple pathways resulting in poor reproducibility78”
From Organoids (2022) · cites this paper as [78]
The Abstract links reliance on recombinant growth factors to cost and poor reproducibility, and the Discussion says the small-molecule approach addresses cost concerns. The cited paper does not say that growth factors are unstable or that small molecules affect multiple pathways and thereby cause poor reproducibility. (LLM, conf=0.86)
Experimental — accuracy not yet measuredThe 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 [18]
The Phase III Results and Discussion describe a protocol developed using published methods and optimized to produce hepatocyte-like cells (HLCs). The Results section ‘Small-Molecule-Derived HLCs Demonstrate Hepatic Function’ reports CYP3A4 activity and its induction. The paper does not report generating iHLOs (organoids) or say that high CYP3A4 activity was the specific reason for choosing the protocol. Page numbers are unavailable in our copy. (LLM, conf=0.87)
Experimental — accuracy not yet measuredThe 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 [18]
The Results section, “Small-Molecule-Derived HLCs Demonstrate Hepatic Function” (page unavailable), reports higher basal CYP1A2 and CYP3A4 activity in the protocol-derived cells than in undifferentiated controls, as well as inducible activity. However, the paper identifies its products as hepatocyte-like cells (HLCs), not iHLOs, and does not demonstrate that the protocol generates organoids with that activity. (LLM, conf=0.89)
Experimental — accuracy not yet measured
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Cites
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Directed differentiation of human embryonic stem cells into functional hepatic cells†
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“Some recent progress has been made in replacing growth factors for the differentiation of mesoderm and ectoderm (Chambers et al., 2012; Lian et al., 2012), and efforts have been undertaken to find suitable candidates for the production of definitive endoderm (DE), exemplified by the Melton group, who identified IDE1 a…”
BackgroundDifferentiation and Transplantation of Human Embryonic Stem Cell–Derived Hepatocytes
Basma, H., Soto–Gutiérrez, A., Yannam, G. R., Liu, L., Ito, R., Yamamoto, T., Ellis, E. C. S., Carson, S. D., Sato, S., Chen, Y., et al. (2008) · Gastroenterology · doi:10.1053/j.gastro.2008.10.047 · Cited by 509 · cited 1× · see citing works
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BackgroundEfficient generation of hepatocyte-like cells from human induced pluripotent stem cells
Song, Z., Cai, J., Liu, Y., Zhao, D., Yong, J., Duo, S., Song, X., Guo, Y., Zhao, Y., Qin, H., et al. (2009) · Cell Research · doi:10.1038/cr.2009.107 · Cited by 499 · cited 1× · see citing works
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BackgroundTouboul, T., Hannan, N. R. F., Corbineau, S., Martinez, A., Martinet, C., Branchereau, S., Goulinet, S., Strick‐Marchand, H., Pedersen, R. A., Santo, J. P. D., et al. (2009) · Hepatology · doi:10.1002/hep.23506 · Cited by 499 · cited 3× · see citing works
“…growth factor (VEGF), epidermal growth factor (EGF), and bone morphogenetic protein 4 (BMP4) (Agarwal et al., 2008; Brolén et al., 2010; Cai et al., 2007; Chen et al., 2012; Hay et al., 2008; Liu et al., 2010; Si-Tayeb et al., 2010b; Song et al., 2009; Sullivan et al., 2010; Touboul et al., 2010).”
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Generation of Functional Human Hepatic Endoderm From Human Induced Pluripotent Stem Cells
Sullivan, G. J., Hay, D. C., Park, I.-H., Fletcher, J., Hannoun, Z., Payne, C. M., Dalgetty, D., Black, J. R., Ross, J. A., Samuel, K., et al. (2010) · Hepatology · doi:10.1002/hep.23335 · Cited by 445 · cited 10× · see citing works
“We finally settled on a modified formulation of Leibovitz L-15 medium (L-15), which has been described in the literature as a standard method to generate mature hepatocytes (Hay et al., 2008; Sullivan et al., 2010).”
BackgroundMethodInfluentialSelf-citationGeneration of Hepatocyte-Like Cells from Human Embryonic Stem Cells
Rambhatla, L., Chiu, C.-P., Kundu, P., Peng, Y., & Carpenter, M. K. (2003) · Cell Transplantation · doi:10.3727/000000003783985179 · Cited by 437 · cited 1× · see citing works
“There are numerous reports of the utility of the small molecule DMSO in stem cell differentiation and specifically in the generation of hepatic progenitors (Hay et al., 2008; Rambhatla et al., 2003; Soto-Gutierrez et al., 2006; Sullivan et al., 2010).”
BackgroundHay, D. C., Fletcher, J. M., Payne, C. M., Terrace, J. D., Gallagher, R., Snoeys, J., Black, J. R., Wojtacha, D., Samuel, K., Hannoun, Z., et al. (2008) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0806522105 · Cited by 434 · cited 16× · see citing works
“We finally settled on a modified formulation of Leibovitz L-15 medium (L-15), which has been described in the literature as a standard method to generate mature hepatocytes (Hay et al., 2008; Sullivan et al., 2010).”
BackgroundMethodInfluentialDifferentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro
Baharvand, H., Hashemi, S. M., Kazemi Ashtiani, S., & Farrokhi, A. (2006) · The International Journal of Developmental Biology · doi:10.1387/ijdb.052072hb · Cited by 429 · cited 1× · see citing works
“Early protocols for the generation of hepatocytes from pluripotent cells relied on the use of embryoid body formation (Baharvand et al., 2006; Basma et al., 2009; Imamura et al., 2004).”
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BackgroundEfficient Differentiation of Functional Hepatocytes from Human Embryonic Stem Cells
Agarwal, S., Holton, K., & Lanza, R. (2008) · Stem Cells · doi:10.1634/stemcells.2007-1102 · Cited by 402 · cited 2× · see citing works
“…growth factor 4 (FGF4), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and bone morphogenetic protein 4 (BMP4) (Agarwal et al., 2008; Brolén et al., 2010; Cai et al., 2007; Chen et al., 2012; Hay et al., 2008; Liu et al., 2010; Si-Tayeb et al., 2010b; Song et…”
BackgroundHepatocyte migration during liver development requires Prox1
Sosa-Pineda, B., Wigle, J. T., & Oliver, G. (2000) · Nature Genetics · doi:10.1038/76996 · Cited by 381 · cited 2× · see citing works
“4 j 939–952 j May 12, 2015 j a2015 The Autho there was an increase in expression of PROX1 and TBX3, which are believed to interact in vivo to promote themigration and proliferation of hepatoblasts from the primary liver bud (Sosa-Pineda et al., 2000; Lüdtke et al., 2009).”
BackgroundMethodGastrulation in the mouse: The role of the homeobox gene goosecoid
Blum, M., Gaunt, S. J., Cho, K. W. Y., Steinbeißer, H., Blumberg, B., Bittner, D., & Robertis, E. M. D. (1992) · Cell · doi:10.1016/0092-8674(92)90632-m · Cited by 376 · cited 3× · see citing works
“We monitored developmentally relevant markers of primitive streak (T, MIXL1, GSC, and FOXA2), mesendoderm (T and FOXA2), and definitive endoderm (HHEX, CER1, SOX17, and FOXA2).”
BackgroundProduction of hepatocyte-like cells from human pluripotent stem cells
Hannan, N. R. F., Segeritz, C., Touboul, T., & Vallier, L. (2013) · Nature Protocols · doi:10.1038/nprot.2012.153 · Cited by 327 · see citing works
Chen, Y.-F., Tseng, C.-Y., Wang, H.-W., Kuo, H.-C., Yang, V. W., & Lee, O. K. (2012) · Hepatology · doi:10.1002/hep.24790 · Cited by 291 · cited 1× · see citing works
“…growth factor (VEGF), epidermal growth factor (EGF), and bone morphogenetic protein 4 (BMP4) (Agarwal et al., 2008; Brolén et al., 2010; Cai et al., 2007; Chen et al., 2012; Hay et al., 2008; Liu et al., 2010; Si-Tayeb et al., 2010b; Song et al., 2009; Sullivan et al., 2010; Touboul et al., 2010).”
BackgroundGeneration of Endoderm-Derived Human Induced Pluripotent Stem Cells From Primary Hepatocytes
Liu, H., Ye, Z., Kim, Y., Sharkis, S. J., & Jang, Y.-Y. (2010) · Hepatology · doi:10.1002/hep.23626 · Cited by 247 · cited 3× · see citing works
“…growth factor (VEGF), epidermal growth factor (EGF), and bone morphogenetic protein 4 (BMP4) (Agarwal et al., 2008; Brolén et al., 2010; Cai et al., 2007; Chen et al., 2012; Hay et al., 2008; Liu et al., 2010; Si-Tayeb et al., 2010b; Song et al., 2009; Sullivan et al., 2010; Touboul et al., 2010).”
BackgroundFrom fertilization to gastrulation: axis formation in the mouse embryo
Lu, C. C., Brennan, J., & Robertson, E. J. (2001) · Current Opinion in Genetics & Development · doi:10.1016/s0959-437x(00)00208-2 · Cited by 238 · cited 1× · see citing works
“We observed a rapid upregulation of NODAL within 4 hr of exposure to CHIR99021, which is indicative of a transition toward a PS population (Figure 3) (Lu et al., 2001).”
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“Phase I of differentiation consists of a 24-hr treatment with RPMI-B27 ± insulin (RPMI 1640 GlutaMAX + B27 supplement, both from Life Technologies) plus 3–4 mM CHIR99021 (Stemgent), followed by a 24-hr treatment with RPMI-B27 alone.”
BackgroundInfluentialSoto–Gutiérrez, A., Navarro–Álvarez, N., Zhao, D., Rivas‐Carrillo, J. D., LEBKOWSKI, J. S., Tanaka, N., Fox, I. J., & Kobayashi, N. (2007) · Nature Protocols · doi:10.1038/nprot.2007.18 · Cited by 132 · cited 2× · see citing works
“The use of DMSO to drive DE toward a hepatic fate is well established in the field (Hay et al., 2008; Soto-Gutiérrez et al., 2007; Sullivan et al., 2010), and we show here that it is equally successful when employed following CHIR99021 treatment as with activin A and Wnt3a.”
Brolén, G., Sivertsson, L., Björquist, P., Eriksson, G., Ek, M., Semb, H., Johansson, I., Andersson, T. B., Ingelman‐Sundberg, M., & Heins, N. (2009) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2009.11.007 · Cited by 120 · cited 3× · see citing works
“…4 (FGF4), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and bone morphogenetic protein 4 (BMP4) (Agarwal et al., 2008; Brolén et al., 2010; Cai et al., 2007; Chen et al., 2012; Hay et al., 2008; Liu et al., 2010; Si-Tayeb et al., 2010b; Song et al., 2009; Sullivan et…”
BackgroundLüdtke, T. H. ‐., Christoffels, V. M., Petry, M., & Kispert, A. (2009) · Hepatology · doi:10.1002/hep.22700 · Cited by 117 · cited 1× · see citing works
“…of PROX1 and TBX3, which are believed to interact in vivo to promote themigration and proliferation of hepatoblasts from the primary liver bud (Sosa-Pineda et al., 2000; Lüdtke et al., 2009). rs The human embryonic stem cell line H1 was differentiated in RPMI-B27 supplemented with 3 mM CHIR99021.”
MethodAccurate Prediction of Drug-Induced Liver Injury Using Stem Cell-Derived Populations
Szkolnicka, D., Farnworth, S. L., Lucendo‐Villarin, B., Storck, C., Zhou, W., Iredale, J. P., Flint, O. P., & Hay, D. C. (2013) · Stem Cells Translational Medicine · doi:10.5966/sctm.2013-0146 · Cited by 105 · cited 1× · see citing works
“Initially, phase III was optimized using the commercially available base mediumHepatoZYME that has been shown to provide supportive conditions for hepatocyte maturation (Szkolnicka et al., 2014).”
MethodAsahina, K., Fujimori, H., Shimizu‐Saito, K., Kumashiro, Y., Okamura, K., Tanaka, Y., Teramoto, K., Arii, S., & Teraoka, H. (2004) · Genes to Cells · doi:10.1111/j.1365-2443.2004.00809.x · Cited by 103 · cited 1× · see citing works
“To corroborate that the source of the HLCs is from DE rather than from yolk sac, we demonstrated that the DE origin marker CYP7A1 was expressed (Asahina et al., 2004) (Figure 6F).”
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“The rationale to examine GSK-3 rs inhibition as a route to producing DE has a basis in the observations of Lickert and colleagues (Engert et al., 2013), who demonstrated that Wnt signaling regulates SOX17 expression.”
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“Early protocols for the generation of hepatocytes from pluripotent cells relied on the use of embryoid body formation (Baharvand et al., 2006; Basma et al., 2009; Imamura et al., 2004).”
BackgroundTan, J. Y., Sriram, G., Rufaihah, A. J., Neoh, K. G., & Cao, T. (2013) · Stem Cells and Development · doi:10.1089/scd.2012.0590 · Cited by 90 · cited 14× · see citing works
“Recent studies have also demonstrated the utility of other small molecules in priming pluripotent cells for differentiation toward endoderm (Tahamtani et al., 2013) and mesoderm (Tan et al., 2013).”
BackgroundInfluentialNakanishi, M., Kurisaki, A., Hayashi, Y., Warashina, M., Ishiura, S., Furue, M. K., & Asashima, M. (2008) · The FASEB Journal · doi:10.1096/fj.08-111203 · Cited by 86 · cited 2× · see citing works
“A number of publications have demonstrated that Wnt signaling is required to specify DE (Engert et al., 2013; Nakanishi et al., 2009), and we have found that a pulse of GSK-3 inhibition is sufficient to produce Cell Reports j Vol. 4 j 939–952 j May 12, 2015 j ª2015 The Authors 939 (A) The normal…”
BackgroundSineva, G. S., & Pospelov, V. A. (2010) · Biology of the Cell · doi:10.1042/bc20100016 · Cited by 67 · cited 1× · see citing works
“…BIO and CHIR99021 are potent pharmacological GSK3-specific inhibitors that result in activation of the Wnt signaling pathway (Sato et al., 2004; Sineva and Pospelov, 2010), so we next assessed the ability of the protein Wnt3a alone to drive differentiation toward DE. Figure 2C demonstrates…”
BackgroundSoto–Gutiérrez, A., Navarro–Álvarez, N., Rivas‐Carrillo, J. D., Chen, Y., Yamatsuji, T., Tanaka, N., & Kobayashi, N. (2006) · Cell Transplantation · doi:10.3727/000000006783981945 · Cited by 57 · cited 1× · see citing works
“There are numerous reports of the utility of the small molecule DMSO in stem cell differentiation and specifically in the generation of hepatic progenitors (Hay et al., 2008; Rambhatla et al., 2003; Soto-Gutierrez et al., 2006; Sullivan et al., 2010).”
BackgroundEvaluation of Metabolically Stabilized Angiotensin IV Analogs as Procognitive/Antidementia Agents
McCoy, A. T., Benoist, C. C., Wright, J. W., Kawas, L. H., Bule-Ghogare, J. M., Zhu, M., Appleyard, S. M., Wayman, G. A., & Harding, J. W. (2012) · Journal of Pharmacology and Experimental Therapeutics · doi:10.1124/jpet.112.199497 · Cited by 54 · cited 3× · see citing works
“We identified a potent HGF mimetic called N-hexanoic-Tyr, Ile-(6) aminohexanoic amide, or dihexa, originally developed as an anti-dementia drug (McCoy et al., 2013).”
BackgroundInfluentialDerivation of 30 human embryonic stem cell lines—improving the quality
Ström, S., Holm, F., Bergström, R., Strömberg, A., & Hovatta, O. (2010) · In vitro Cellular & Developmental Biology-Animal · doi:10.1007/s11626-010-9308-0 · Cited by 53 · cited 1× · see citing works
“H1 hESCs (WiCell) and 207 hESCs (Ström et al., 2010) and Detroit 551 (ATCC CCL-110) hiPSCs were maintained at 37 C/5% CO2 in feeder-free conditions using growth-factor-reduced Matrigel (Sigma-Aldrich) and E8 Medium (Life Technologies), with routine passaging performed at a 1:3 ratio using 0.5 mM…”
MethodInfluentialThe Making of the Liver: Competence in the Foregut Endoderm and Induction of Liver-Specific Genes
Kaestner, K. H. (2005) · Cell Cycle · doi:10.4161/cc.4.9.2033 · Cited by 46 · cited 2× · see citing works
“See also Figure S2. progenitors, we observed maintenance of FOXA2 levels and an induction of GATA4, both of which are known to be pioneer factors critical to promote the hepatic gene expression program (Kaestner, 2005).”
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“Following studies of the utility of GSK-3 inhibition in priming pluripotent cells for endodermal differentiation (Tahamtani et al., 2013) and reports that Wnt/b-catenin signaling regulates SOX17 expression and is essential for endoderm formation (Engert et al., 2013), we set out to establish…”
BackgroundGeneration of Functional Hepatic Cells from Pluripotent Stem Cells
Han, S. (2012) · Journal of Stem Cell Research & Therapy · doi:10.4172/2157-7633.s10-008 · Cited by 23 · cited 6× · see citing works
“However, HepatoZYME is a proprietary reagent and contains the growth factor EGF and thus was unsuitable for this application (Garcia et al., 2001).”
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What are the limits to cell plasticity?
Taylor, J., Wilmut, I., & Sullivan, G. J. (2010) · Cell Research · doi:10.1038/cr.2010.59 · Cited by 6 · cited 1× · see citing works
“Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) offer a potentially limitless source of cells for industrial and clinical translation, and the everadvancing field of cellular reprogramming has redefined the limits of cell plasticity (Taylor et al., 2010).”
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Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes
Cameron, K., Tan, R., Schmidt‐Heck, W., Campos, G., Lyall, M. J., Wang, Y., Lucendo‐Villarin, B., Szkolnicka, D., Bates, N., Kimber, S. J., et al. (2015) · Stem Cell Reports · doi:10.1016/j.stemcr.2015.10.016 · Cited by 149 · see citing works
“Other studies have used small molecules to replace growth factors to drive down the cost of the method (Hay et al., 2007, 2008; Siller et al., 2015); however, those publishedmethods both still rely upon the use of undefined components, highlighting the need to develop defined systems.”
MethodLiver Organoids: Recent Developments, Limitations and Potential
Harrison, S. P., Baumgarten, S., Verma, R., Lunov, O., Dejneka, A., & Sullivan, G. J. (2021) · Frontiers in Medicine · doi:10.3389/fmed.2021.574047 · Cited by 141 · see citing works
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Fernández-Checa, J. C., Bagnaninchi, P., Ye, H., Sancho‐Bru, P., Falcón‐Pérez, J. M., Royo, F., García‐Ruiz, C., Konu, Ö., Miranda, J. P., Lunov, O., et al. (2021) · Journal of Hepatology · doi:10.1016/j.jhep.2021.06.021 · Cited by 129 · see citing works
Self-citationCurrent progress in hepatic tissue regeneration by tissue engineering
Hosseini, V., Maroufi, N. F., Saghati, S., Asadi, N., Darabi, M., Ahmad, S. N. S., Hosseinkhani, H., & Rahbarghazi, R. (2019) · Journal of Translational Medicine · doi:10.1186/s12967-019-02137-6 · Cited by 122 · see citing works
Scalable production of tissue-like vascularized liver organoids from human PSCs
Harrison, S. P., Siller, R., Tanaka, Y., Chollet, M. E., de la Morena-Barrio, M. E., Xiang, Y., Patterson, B., Andersen, E., Bravo-Pérez, C., Kempf, H., et al. (2023) · Experimental & Molecular Medicine · doi:10.1038/s12276-023-01074-1 · Cited by 119 · see citing works
Self-citationBiotechnology Challenges to In Vitro Maturation of Hepatic Stem Cells
Chen, C., Soto–Gutiérrez, A., Baptista, P. M., & Spee, B. (2018) · Gastroenterology · doi:10.1053/j.gastro.2018.01.066 · Cited by 103 · see citing works
Du, C., Feng, Y., Qiu, D., Xu, Y., Pang, M., Cai, N., Xiang, A. P., & Zhang, Q. (2018) · Stem Cell Research & Therapy · doi:10.1186/s13287-018-0794-4 · Cited by 106 · see citing works
“In contrast to published protocols using RPMI 1640 and B-27 Supplement as the basal medium [34], we also changed the basal medium to RPMI 1640 and B-27 Supplement Minus Insulin to improve the definitive endoderm generation efficiency.”
BackgroundResultInfluentialThe potential of induced pluripotent stem cell derived hepatocytes
Hannoun, Z., Steichen, C., Dianat, N., Weber, A., & Dubart‐Kupperschmitt, A. (2016) · Journal of Hepatology · doi:10.1016/j.jhep.2016.02.025 · Cited by 103 · see citing works
“pluripotent stem cells to hepatocytes [58].”
Halloin, C., Schwanke, K., Löbel, W., Franke, A., Szepes, M., Biswanath, S., Wunderlich, S., Merkert, S., Weber, N., Osten, F., et al. (2019) · Stem Cell Reports · doi:10.1016/j.stemcr.2019.06.004 · Cited by 100 · see citing works
“Notably, chemical WNT pathway stimulators (particularly the GSK3 inhibitor CHIR99021 [CHIR]) or suppressors (including IWP2, IWR1, and Wnt-C59) have also been applied to specify other mesendodermal lineages including hepatocytes (Siller et al., 2015) and skeletal muscle cells (Shelton et al.”
BackgroundComparing in vitro human liver models to in vivo human liver using RNA-Seq
Gupta, R., Schrooders, Y., Hauser, D. A., van Herwijnen, M. H. M., Albrecht, W., ter Braak, B., Brecklinghaus, T., Castell, J. V., Elenschneider, L., Escher, S. E., et al. (2020) · Archives of Toxicology · doi:10.1007/s00204-020-02937-6 · Cited by 94 · see citing works
Laco, F., Woo, T. L., Zhong, Q., Szmyd, R., Ting, S., Khan, F. J., Chai, C. L. L., Reuveny, S., Chen, A., & Oh, S. (2018) · Stem Cell Reports · doi:10.1016/j.stemcr.2018.03.023 · Cited by 96 · see citing works
“A low CHIR concentration has been shown to lead to endoderm development, which can induce hepatocyte differentiation (Siller et al., 2015).”
BackgroundHuang, J., Guo, X., Li, W., & Zhang, H. (2017) · Scientific Reports · doi:10.1038/srep40716 · Cited by 95 · see citing works
Small molecule–driven direct conversion of human pluripotent stem cells into functional osteoblasts
Kang, H., Shih, Y.-R. V., Nakasaki, M., Kabra, H., & Varghese, S. (2016) · Science Advances · doi:10.1126/sciadv.1600691 · Cited by 80 · see citing works
Stem Cell-Based Therapies for Liver Diseases: An Overview and Update
Wang, J., Sun, M., Liu, W., Li, Y., & Li, M. (2019) · Tissue Engineering and Regenerative Medicine · doi:10.1007/s13770-019-00178-y · Cited by 76 · see citing works
“Most of them apply growth factors and cytokines such as activin A, Wnt3a, hepatocyte growth factor (HGF), oncostatin M (OsM) to induce liver development in vitro [76, 77].”
BackgroundHuman Pluripotent Stem Cell-Derived Endoderm for Modeling Development and Clinical Applications
Yiangou, L., Ross, A., Goh, K. J., & Vallier, L. (2018) · Cell stem cell · doi:10.1016/j.stem.2018.03.016 · Cited by 76 · see citing works
“…be involved in liver development and functional maturation (Agarwal et al., 2008; Brolén et al., 2010; Cai et al., 2007; Chen et al., 2012; Hannan et al., 2013b;Mallanna and Duncan, 2013; Si-Tayeb et al., 2010; Siller et al., 2015; Song et al., 2009; Sullivan et al., 2010; Touboul et al., 2010).”
Therapeutic Potential of Differentiated Mesenchymal Stem Cells for Treatment of Osteoarthritis
Ham, O., Lee, C. Y., Kim, R., Lee, J., Oh, S., Lee, M., Kim, J., Hwang, K., Maeng, L.-S., & Chang, W. (2015) · International Journal of Molecular Sciences · doi:10.3390/ijms160714961 · Cited by 78 · see citing works
Direct On-Chip Differentiation of Intestinal Tubules from Induced Pluripotent Stem Cells
Naumovska, E., Aalderink, G., Valencia, C. W., Kosim, K., Nicolas, A., Brown, S., Vulto, P., Erdmann, K. S., & Kurek, D. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21144964 · Cited by 72 · see citing works
“GSK-3 inhibition by CHIR99021 alone is sufficient for the generation of SOX17 and FOXA2 positive cells [19,47].”
BackgroundSelf-citationLiver cell therapy: is this the end of the beginning?
Alwahsh, S. M., Rashidi, H., & Hay, D. C. (2017) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-017-2713-8 · Cited by 70 · see citing works
“have devised a growth factor-free protocol that relies on small molecules to efficiently differentiate human PSCs toward a hepatic phenotype [103], however, their approach still relied on fetal bovine serum and Matrigel, limiting GMP-grade.”
BackgroundMoskot, M., Jakóbkiewicz‐Banecka, J., Kłoska, A., Piotrowska, E., Narajczyk, M., & Gabig‐Cimińska, M. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20020304 · Cited by 62 · see citing works
“5–5% v/v, or even 10% v/v, it is frequently applied to induce differentiation of various cell lines [34], including fibroblasts [31,32].”
BackgroundLiver diseases in the dish: iPSC and organoids as a new approach to modeling liver diseases
Fiorotto, R., Amenduni, M., Mariotti, V., Fabris, L., Spirlı̀, C., & Strazzabosco, M. (2018) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/j.bbadis.2018.08.038 · Cited by 63 · see citing works
Hepatic differentiation of human iPSCs in different 3D models: A comparative study
Meier, F., Freyer, N., Brzeszczyńska, J., spel, F. K. ⁄., Armstrong, L., Lako, M., Greuel, S., Damm, G., Ludwig-Schwellinger, E., Deschl, U., et al. (2017) · International Journal of Molecular Medicine · doi:10.3892/ijmm.2017.3190 · Cited by 59 · see citing works
“The generated HLCs demonstrate some characteristics of hepatocytes, such as susceptibility to hepatitis C virus infection (13), secretion of hepatic proteins (14,15) and activity of metabolic enzymes (16,17).”
Disease‐specific phenotypes in iPSC‐derived neural stem cells with POLG mutations
Liang, K. X., Kristiansen, C. K., Mostafavi, S., Vatne, G. H., Zantingh, G. A., Kianian, A., Tzoulis, C., Høyland, L. E., Ziegler, M., Pérez, R., et al. (2020) · EMBO Molecular Medicine · doi:10.15252/emmm.202012146 · Cited by 60 · see citing works
“Hepatocyte differentiation from iPSCs was optimized for patient and control lines using a previously established approach (Siller et al, 2015, 2016; Siller & Sullivan, 2017).”
MethodSelf-citationBiomaterials used in stem cell therapy for spinal cord injury
Higuchi, A., Kumar, S. S., Benelli, G., Ling, Q., Li, H.-F., Alarfaj, A. A., Munusamy, M. A., Sung, T., Chang, Y., & Murugan, K. (2019) · Progress in Materials Science · doi:10.1016/j.pmatsci.2019.02.002 · Cited by 59 · see citing works
Generation of heart-forming organoids from human pluripotent stem cells
Drakhlis, L., Biswanath, S., & Zweigerdt, R. (2021) · Nature Protocols · doi:10.1038/s41596-021-00629-8 · Cited by 58 · see citing works
Cox, C. R., Lynch, S., Goldring, C. E., & Sharma, P. (2020) · Frontiers in Medical Technology · doi:10.3389/fmedt.2020.611913 · Cited by 57 · see citing works
“This includes the use of growth factors and small molecules (61, 154, 155), small molecules alone (59, 156), or transcription factors (157).”
BackgroundGraffmann, N., Scherer, B., & Adjaye, J. (2022) · Stem Cell Research · doi:10.1016/j.scr.2022.102763 · Cited by 54 · see citing works
Differentiation of hepatocyte-like cells from human pluripotent stem cells using small molecules
Asumda, F. Z., Hatzistergos, K. E., Dykxhoorn, D. M., Jakubski, S., Edwards, J. S., Thomas, E., & Schiff, E. R. (2018) · Differentiation · doi:10.1016/j.diff.2018.03.002 · Cited by 52 · see citing works
“Accordingly, here we tested the efficacy of a previously published (Siller et al., 2015; Mathapati et al., 2016) small-molecule based differentiation protocol for deriving HLCs with a mature phenotype from hPSCs.”
BackgroundMethodInfluentialBalafkan, N., Mostafavi, S., Schubert, M., Siller, R., Liang, K. X., Sullivan, G. J., & Bindoff, L. A. (2020) · Scientific Reports · doi:10.1038/s41598-020-73656-2 · Cited by 52 · see citing works
Self-citationPluripotent stem cells to hepatocytes, the journey so far
Palakkan, A. A., Nanda, J., & Ross, J. A. (2017) · Biomedical Reports · doi:10.3892/br.2017.867 · Cited by 49 · see citing works
“3 μM CHIR99021(mimic 1 FOXA2, SOX17, (55)”
Myotubes derived from human-induced pluripotent stem cells mirror in vivo insulin resistance
Iovino, S., Burkart, A., Warren, L., Patti, M., & Kahn, C. R. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1525665113 · Cited by 48 · see citing works
Fetal liver development and implications for liver disease pathogenesis
Lotto, J., Stephan, T. L., & Hoodless, P. A. (2023) · Nature Reviews Gastroenterology & Hepatology · doi:10.1038/s41575-023-00775-2 · Cited by 47 · see citing works
Small‐Molecule‐Directed Hepatocyte‐Like Cell Differentiation of Human Pluripotent Stem Cells
Mathapati, S., Siller, R., Impellizzeri, A. A. R., Lycke, M., Vegheim, K., Almaas, R., & Sullivan, G. J. (2016) · Current Protocols in Stem Cell Biology · doi:10.1002/cpsc.13 · Cited by 47 · see citing works
Self-citationGurevich, I., Burton, S., Munn, C. G., Ohshima, M., Goedland, M., Czysz, K., & Rajesh, D. (2020) · Biology Open · doi:10.1242/bio.055087 · Cited by 47 · see citing works
“The protocol evolved from several published (Mallanna and Duncan, 2013; Peters et al., 2016; Siller et al., 2015; Takayama et al., 2012) hepatocyte differentiation protocols (Fig.”
Yan, Y., Bejoy, J., Xia, J., Guan, J., Zhou, Y., & Li, Y. (2016) · Acta Biomaterialia · doi:10.1016/j.actbio.2016.06.027 · Cited by 45 · see citing works
Hepatocyte-like cells derived from induced pluripotent stem cells
Roy‐Chowdhury, N., Wang, X., Guha, C., & Roy‐Chowdhury, J. (2016) · Hepatology International · doi:10.1007/s12072-016-9757-y · Cited by 45 · see citing works
Siller, R., Naumovska, E., Mathapati, S., Lycke, M., Greenhough, S., & Sullivan, G. (2016) · Scientific Reports · doi:10.1038/srep37178 · Cited by 43 · see citing works
Self-citationDevelopment of organoid-based drug metabolism model
Park, E., Kim, H. K., Jee, J., Hahn, S., Jeong, S., & Yoo, J. (2019) · Toxicology and Applied Pharmacology · doi:10.1016/j.taap.2019.114790 · Cited by 44 · see citing works
Szkolnicka, D., & Hay, D. C. (2016) · Stem Cells · doi:10.1002/stem.2368 · Cited by 42 · see citing works
Advancements in stem cell-derived hepatocyte-like cell models for hepatotoxicity testing
Jin, M., Xiao, Y., Liao, W., Chen, Q., Yang, W., Li, Y., Li, S., Gao, Y., Peng, Q., & Zhou, S. (2021) · Stem Cell Research & Therapy · doi:10.1186/s13287-021-02152-9 · Cited by 41 · see citing works
“Researchers have shown that the use of small molecules [42, 43] regulating the extracellular nutrient level [44, 45], inducing overexpression of hepatic transcription factors [46, 47], and manipulating miRNA expression [48, 49] represents powerful methods for the efficient generation of metabolically functional hepato…”
BackgroundGao, X., Li, R., Cahan, P., Zhao, Y., Yourick, J. J., & Sprando, R. L. (2020) · Stem Cell Research & Therapy · doi:10.1186/s13287-020-01914-1 · Cited by 40 · see citing works
Li, Q., Lin, H., Du, Q., Liu, K., Wang, O., Evans, C. S., Christian, H., Zhang, C., & Lei, Y. (2018) · Biofabrication · doi:10.1088/1758-5090/aaa6b5 · Cited by 37 · see citing works
Oxygen drives hepatocyte differentiation and phenotype stability in liver cell lines
van Wenum, M., Adam, A. A. A., van der Mark, V. A., Chang, J., Wildenberg, M. E., Hendriks, E. J., Jongejan, A., Moerland, P. D., van Gulik, T. M., Elferink, R. P. J. O., et al. (2018) · Journal of Cell Communication and Signaling · doi:10.1007/s12079-018-0456-4 · Cited by 38 · see citing works
“…(Huch et al. 2015), and of ways to influence these in vitro through co-culturing (Kidambi et al. 2009; Takebe et al. 2013), culture platforms (vanWenum et al. 2014), small molecules (Huch et al. 2015; Siller et al. 2015) and extracellular matrix constructs (Park et al. 2016; Dunn et al. 1989).”
BackgroundStem-cell derived hepatocyte-like cells for the assessment of drug-induced liver injury
Donato, M. T., & Tolosa, L. (2019) · Differentiation · doi:10.1016/j.diff.2019.02.004 · Cited by 37 · see citing works
Parmentier, C., Couttet, P., Wolf, A., Zaccharias, T., Heyd, B., Bachellier, P., Uteng, M., & Richert, L. (2017) · Archives of Toxicology · doi:10.1007/s00204-017-1930-0 · Cited by 36 · see citing works
“The measure of mRNA signatures of hepatocyte phase I, II and II differentiation stages (Siller et al. 2015) across the five PHH cultures showed a predominance of fully matured hepatocytes , which was, moreover, not affected by the 14 days of 2D-sandwich culture.”
BackgroundMethodSmall molecule-induced cellular conversion
De, D., Halder, D., Shin, I., & Kim, K. K. (2017) · Chemical Society Reviews · doi:10.1039/c7cs00330g · Cited by 36 · see citing works
Tricot, T., Verfaillie, C. M., & Kumar, M. (2022) · Cells · doi:10.3390/cells11030442 · Cited by 35 · see citing works
“To enhance fating of HLCs to the level of PHHs, adapted differentiation protocols have included the addition of small molecules that influence different signaling pathways believed to play a role in PHH development, during the differentiation protocol [65,81,82], the use of medium that has optimized nutrient levels [7…”
BackgroundGeneration of human pluripotent stem cell-derived hepatocyte-like cells for drug toxicity screening
Takayama, K., & Mizuguchi, H. (2016) · Drug Metabolism and Pharmacokinetics · doi:10.1016/j.dmpk.2016.10.408 · Cited by 33 · see citing works
Govarthanan, K., Vidyasekar, P., Gupta, P. K., Lenka, N., & Verma, R. S. (2020) · Cytotherapy · doi:10.1016/j.jcyt.2019.12.007 · Cited by 33 · see citing works
Varghese, D. S., Alawathugoda, T. T., & Ansari, S. A. (2019) · Stem Cells International · doi:10.1155/2019/5968236 · Cited by 32 · see citing works
“Both methods of DE induction from hESCs have been used in the previous studies [9, 12].”
BackgroundMethodElectromembrane Extraction and Mass Spectrometry for Liver Organoid Drug Metabolism Studies
Skottvoll, F. S., Hansen, F. A., Harrison, S. P., Boger, I. C. S., Mrsa, A., Restan, M. S., Stein, M., Lundanes, E., Pedersen‐Bjergaard, S., Aizenshtadt, A., et al. (2021) · Analytical Chemistry · doi:10.1021/acs.analchem.0c05082 · Cited by 32 · see citing works
Self-citationKim, J., Kim, Y., Choi, J., Jung, H., Lee, K.-J., Kang, J., Park, N., Rim, Y. A., Nam, Y., & Ju, J. H. (2018) · Stem Cell Research & Therapy · doi:10.1186/s13287-018-1100-1 · Cited by 32 · see citing works
Cotovio, J. P., & Fernandes, T. G. (2020) · Bioengineering · doi:10.3390/bioengineering7020036 · Cited by 30 · see citing works
“, 2015 RPMI+B27 – DMEM+SR+DMSO – L15 CHIR – Dihexa/DEX [48]”
BackgroundHydrogel-Based Strategies for Liver Tissue Engineering
Zhang, Y., Li, L., Dong, L., Cheng, Y., Huang, X., Xue, B., Jiang, C., Cao, Y., & Yang, J. (2024) · Chem & Bio Engineering · doi:10.1021/cbe.4c00079 · Cited by 31 · see citing works
Li, Z., Wu, J., Wang, L., Han, W., Yu, J., Liu, X., Wang, Y., Zhang, Y., Feng, G., Li, W., et al. (2019) · Cell Death and Disease · doi:10.1038/s41419-019-1967-5 · Cited by 30 · see citing works
Raasch, M., Fritsche, E., Kurtz, A., Bauer, M. W., & Mosig, A. S. (2018) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2018.06.008 · Cited by 30 · see citing works
Gheibi, S., Singh, T., da Cunha, J. P. D., Fex, M., & Mulder, H. (2020) · Cells · doi:10.3390/cells9112465 · Cited by 22 · see citing works
“Expands the PDX1+ PPs and promotes endocrine cell fate [155]”
BackgroundFarzaneh, Z., Najarasl, M., Abbasalizadeh, S., Vosough, M., & Baharvand, H. (2018) · Stem Cells and Development · doi:10.1089/scd.2017.0074 · Cited by 28 · see citing works
“B-27 supplement without insulin or vitamin A have been used to develop a defined and serum-free medium for differentiation of hPSCs to endodermal cells [32,33].”
Advances in generating liver cells from pluripotent stem cells as a tool for modeling liver diseases
Maepa, W., & Ndlovu, H. (2020) · Stem Cells · doi:10.1002/stem.3154 · Cited by 29 · see citing works
Advancements in Disease Modeling and Drug Discovery Using iPSC-Derived Hepatocyte-like Cells
Blaszkiewicz, J., & Duncan, S. A. (2022) · Genes · doi:10.3390/genes13040573 · Cited by 28 · see citing works
Thakur, G., Lee, H.-J., Jeon, R., Lee, S.-L., & Rho, G. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21072388 · Cited by 25 · see citing works
“Small molecules are used to attain many different cell lineages such as osteoblasts [35], neurons [36], cardiomyocytes [37], hepatocytes [38], and pancreaticβ-like cells [19].”
BackgroundXie, Y., Yao, J., Jin, W., Ren, L., & Li, X. (2021) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2021.765980 · Cited by 27 · see citing works
Kempf, H., & Zweigerdt, R. (2017) · Advances in biochemical engineering, biotechnology · doi:10.1007/10_2017_30 · Cited by 26 · see citing works
Technological advancements for the development of stem cell-based models for hepatotoxicity testing
Natale, A., Vanmol, K., Arslan, A., Vlierberghe, S. V., Dubruel, P., Erps, J. V., Hugo Thienpont, A., Buzgo, M., Boeckmans, J., Kock, J. D., et al. (2019) · Archives of Toxicology · doi:10.1007/s00204-019-02465-y · Cited by 25 · see citing works
The Use of Human Pluripotent Stem Cells for Modeling Liver Development and Disease
Heslop, J. A., & Duncan, S. A. (2018) · Hepatology · doi:10.1002/hep.30288 · Cited by 23 · see citing works
Deng, S., Zhao, X., Zhu, Y., Tang, N., Wang, R., Zhang, X., Qu, F., Ho, Y., Lee, W., Chen, J., et al. (2022) · Biofabrication · doi:10.1088/1758-5090/aca79b · Cited by 22 · see citing works
Schöbel, A., Rösch, K., & Herker, E. (2018) · Scientific Reports · doi:10.1038/s41598-018-22243-7 · Cited by 23 · see citing works
Török, G., Erdei, Z., Lilienberg, J., Apáti, Á., & Homolya, L. (2020) · PLoS ONE · doi:10.1371/journal.pone.0227751 · Cited by 22 · see citing works
“These include chemically defined xeno-free media [47,48], different coating substances and 3D culturing compositions [35–39], as well as co-culturing with supporting cells [26], bioprinting of HLCs onto structures mimicking in vivo situation [49].”
BackgroundSignalling pathways and transcriptional regulators orchestrating liver development and cancer
Campbell, S. A., Stephan, T. L., Lotto, J., Cullum, R., Drissler, S., & Hoodless, P. A. (2021) · Development · doi:10.1242/dev.199814 · Cited by 22 · see citing works
Li, Y., Yang, X., Plummer, R., Hayashi, Y., Deng, X.-S., Nie, Y., & Taniguchi, H. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms221910471 · Cited by 24 · see citing works
Parmentier, C., Hendriks, D. F. G., Heyd, B., Bachellier, P., Ingelman‐Sundberg, M., & Richert, L. (2018) · Toxicology Letters · doi:10.1016/j.toxlet.2018.06.1069 · Cited by 22 · see citing works
Tissue-Engineered Bile Ducts for Disease Modeling and Therapy
Wang, Z., Faria, J., Penning, L. C., Masereeuw, R., & Spee, B. (2020) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2020.0283 · Cited by 21 · see citing works
“Because the HNF4 subfamily is exclusively involved in hepatocyte differentiation, 88–90 it is not further discussed in this review.”
Vanhove, J., Pistoni, M., Welters, M., Eggermont, K., Vanslembrouck, V., Helsen, N., Boon, R., Najimi, M., Sokal, E. M., Collas, P., et al. (2016) · Stem Cell Reports · doi:10.1016/j.stemcr.2016.06.013 · Cited by 21 · see citing works
Pisal, R. V., Suchánek, J., Siller, R., Soukup, T., Hrebíková, H., Bezrouk, A., Kunke, D., Mičuda, S., Filip, S., Sullivan, G. J., et al. (2018) · Scientific Reports · doi:10.1038/s41598-018-24421-z · Cited by 20 · see citing works
Self-citationDerivation and applications of human hepatocyte-like cells
Li, S., Huang, S., Zhao, Y., Ding, Y.-J., Ma, D., & Ding, Q. (2019) · World Journal of Stem Cells · doi:10.4252/wjsc.v11.i8.535 · Cited by 20 · see citing works
3D Printed Tooling for Injection Molded Microfluidics
Convery, N., Samardzhieva, I., Stormonth-Darling, J., Harrison, S. P., Sullivan, G. J., & Gadegaard, N. (2021) · Macromolecular Materials and Engineering · doi:10.1002/mame.202100464 · Cited by 20 · see citing works
Self-citationDistinct Mitochondrial Remodeling During Mesoderm Differentiation in a Human-Based Stem Cell Model
Mostafavi, S., Balafkan, N., Pettersen, I. K., Nido, G., Siller, R., Tzoulis, C., Sullivan, G., & Bindoff, L. (2021) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2021.744777 · Cited by 13 · see citing works
Self-citationDifferentiation therapy: Unlocking phenotypic plasticity of hepatocellular carcinoma
Zheng, S., Bian, H., Li, J., Shen, Y., Yang, Y., & Hu, W. (2022) · Critical Reviews in Oncology/Hematology · doi:10.1016/j.critrevonc.2022.103854 · Cited by 20 · see citing works
Liver Bioengineering: Promise, Pitfalls, and Hurdles to Overcome
Acun, A., Oganesyan, R. V., & Uygun, B. E. (2019) · Current Transplantation Reports · doi:10.1007/s40472-019-00236-3 · Cited by 20 · see citing works
Huang, H., Bader, T., & Jin, S. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21165867 · Cited by 19 · see citing works
“For example, enhancing Wingless and Int-1 (WNT) [13], nodal growth differentiation factor (NODAL) [14], and transforming growth factor β (TGF-β) signaling [13] during the generation of definitive endoderm could increase the yield of this lineage.”
BackgroundSong, D., Takahashi, G., Zheng, Y., Matsuo‐Takasaki, M., Li, J., Takami, M., An, Y., Hemmi, Y., Miharada, N., Fujioka, T., et al. (2022) · Human Molecular Genetics · doi:10.1093/hmg/ddac080 · Cited by 19 · see citing works
“A stepwise differentiation protocol was adopted to differentiate iPSCs into hepatocytes in this study, as modified from a previous study (31).”
In Vitro Generated Hepatocyte-Like Cells: A Novel Tool in Regenerative Medicine and Drug Discovery.
Zakikhan, K., Pournasr, B., Massoud, V., & Marjan, N. A. (2017) · PubMed Central · doi:10.22074/cellj.2016.4362 · Cited by 19 · see citing works
“Therefore, numbers of research group have been trying to increase the efficiency of differentiation of iPSCs to HLCs in 2D and 3D with different differentiation protocols (9, 24, 40, 46, 47).”
BackgroundLiang, S., Liang, S., Yin, N., & Faiola, F. (2019) · Ecotoxicology and Environmental Safety · doi:10.1016/j.ecoenv.2019.02.091 · Cited by 19 · see citing works
Is Transforming Stem Cells to Pancreatic Beta Cells Still the Holy Grail for Type 2 Diabetes?
Kahraman, S., Okawa, E. R., & Kulkarni, R. (2016) · Current Diabetes Reports · doi:10.1007/s11892-016-0764-0 · Cited by 16 · see citing works
Addressing Key Questions in Organoid Models: Who, Where, How, and Why?
Gómez‐Álvarez, M., Agustina‐Hernández, M., Francés‐Herrero, E., Rodríguez‐Eguren, A., Bueno-Fernández, C., & Cervelló, I. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms242116014 · Cited by 18 · see citing works
“Thus, one study experimented with combining biologics and small-molecule drugs in organoid cultures and obtained positive results [138].”
BackgroundHuman iPSC-derived liver co-culture spheroids to model liver fibrosis
Cools, L., Dastjerd, M. K., Smout, A., Merens, V., Yang, Y., Reynaert, H., Messaoudi, N., Smet, V. D., Kumar, M., Verhulst, S., et al. (2024) · Biofabrication · doi:10.1088/1758-5090/ad5766 · Cited by 18 · see citing works
“Previous studies have already shown that growth factors can be (partly) replaced by small molecules, leading to successful differentiation of hepatocytes (Siller et al 2015, Pan et al 2022, Vanmarcke et al 2023).”
BackgroundGuo, X., Li, W., Ma, M., Lü, X., & Zhang, H. (2017) · Journal of Cellular and Molecular Medicine · doi:10.1111/jcmm.13195 · Cited by 18 · see citing works
Recent advances in lineage differentiation from stem cells: hurdles and opportunities?
Terryn, J., Tricot, T., Gajjar, M. K., & Verfaillie, C. M. (2018) · F1000Research · doi:10.12688/f1000research.12596.1 · Cited by 18 · see citing works
“Similar examples can be found in the literature for other cell types, including hepatic, neuronal, or cardiac progeny 36,83,84 .”
ResultRaju, R., Chau, D., Notelaers, T., Myers, C. L., Verfaillie, C. M., & Hu, W. (2018) · Stem Cells and Development · doi:10.1089/scd.2017.0270 · Cited by 18 · see citing works
Advancing hepatotoxicity assessment: current advances and future directions
Kim, Y., Kim, Y., & Kim, Y. (2025) · Toxicological Research · doi:10.1007/s43188-025-00289-w · Cited by 20 · see citing works
The translational potential of human induced pluripotent stem cells for clinical neurology
Devine, H. E., & Patani, R. (2016) · Cell Biology and Toxicology · doi:10.1007/s10565-016-9372-7 · Cited by 17 · see citing works
“Hepatocyte-like cells have been differentiated from hiPSCs with success by several groups (Siller et al. 2015).”
BackgroundModeling Inborn Errors of Hepatic Metabolism Using Induced Pluripotent Stem Cells
Pournasr, B., & Duncan, S. A. (2017) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.117.309199 · Cited by 17 · see citing works
Rapid Screening of the Endodermal Differentiation Potential of Human Pluripotent Stem Cells
Siller, R., & Sullivan, G. J. (2017) · Current Protocols in Stem Cell Biology · doi:10.1002/cpsc.36 · Cited by 17 · see citing works
Self-citationAutophagy modulates cell fate decisions during lineage commitment
Sharma, K., Asp, N., Harrison, S. P., Siller, R., Baumgarten, S., Gupta, S., Chollet, M. E., Andersen, E. W., Sullivan, G. J., & Simonsen, A. (2021) · Autophagy · doi:10.1080/15548627.2021.2008691 · Cited by 16 · see citing works
“To efficiently differentiate hiPSCs toward DE, we took advantage of a growth-factor-free protocol that employs the GSK3 (glycogen synthase kinase 3) inhibitor CHIR99021 (WNT agonist) [28,40,41].”
MethodResultInfluentialSelf-citationKalantarMotamedi, Y., Peymani, M., Baharvand, H., Nasr‐Esfahani, M. H., & Bender, A. (2016) · Cell Death Discovery · doi:10.1038/cddiscovery.2016.7 · Cited by 16 · see citing works
Reprogramming cell fate with artificial transcription factors
Heiderscheit, E. A., Eguchi, A., Spurgat, M. C., & Ansari, A. Z. (2018) · FEBS Letters · doi:10.1002/1873-3468.12993 · Cited by 18 · see citing works
Apáti, Á., Varga, N., Berecz, T., Erdei, Z., Homolya, L., & Sarkadi, B. (2018) · Expert Opinion on Drug Metabolism & Toxicology · doi:10.1080/17425255.2019.1558207 · Cited by 16 · see citing works
- Authors
- Siller, R., Greenhough, S., Naumovska, E., & Sullivan, G. J.
- Affiliations
- University of Oslo (NO)
- Oslo University Hospital (NO)
- Corresponding author
- Sullivan, Gareth J.
- Venue
- Stem Cell Reports
- Publisher
- Elsevier BV
- Published
- 2015-05
- Volume
- 4
- Issue
- 5
- Pages
- 939-952
- ISSN
- 2213-6711 (ISSN-L)
- PMC
- PMC4437467
- OpenAlex
- W2012281335
- Cited by
- 203
- References
- 48
- Citations by year
- 2026: 8 · 2025: 9 · 2024: 11 · 2023: 13 · 2022: 19 · 2021: 30 · 2020: 25 · 2019: 15 · 2018: 27 · 2017: 19 · 2016: 19 · 2015: 6 · 2014: 1
- FWCI
- 10.27
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Stem Cell Reports (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- NORA - Norwegian Open Research Archives (submittedVersion, cc-by-nc-nd)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
- Keywords
- hepatocyte differentiation, human pluripotent stem cells, small molecules, human embryonic stem cells, induced pluripotent stem cells, hepatocyte-like cells, glycogen storage, cytochrome P450 activity, hepatic differentiation
- MeSH terms
- Blood Proteins, Cell Differentiation, Cells, Cultured, Dexamethasone, Dimethyl Sulfoxide, Glycogen, Humans, Microscopy, Fluorescence, Oligopeptides, Proto-Oncogene Proteins c-met, Hepatocytes, Glycogen Synthase Kinase 3, Pluripotent Stem Cells, Small Molecule Libraries
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