Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes

Melissa Baxter; Sarah Withey; Sean Harrison; Charis-P Segeritz; Fang Zhang; Rebecca Atkinson-Dell; Cliff Rowe; David Gerrard; Rowena Sison-Young; Roz Jenkins; Joanne Henry; Andrew Berry; Lisa Mohamet; Marie Best; Stephen W. Fenwick; Hassan Malik; Neil R. Kitteringham; Chris E. Goldring; Karen Piper Hanley; Ludovic Vallier; Neil Hanley

doi:10.1016/j.jhep.2014.10.016

Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes

Melissa Baxter, Sarah Withey, Sean Harrison, Charis-P Segeritz, Fang Zhang, Rebecca Atkinson-Dell, Cliff Rowe, David Gerrard, Rowena Sison-Young, Roz Jenkins, Joanne Henry, Andrew Berry, Lisa Mohamet, Marie Best, Stephen W. Fenwick, Hassan Malik, Neil R. Kitteringham, Chris E. Goldring, Karen Piper Hanley, Ludovic VallierNeil Hanley

Division of Diabetes, Endocrinology & Gastroenterology (L5)

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Abstract

BACKGROUND & AIMS: Hepatocyte-like cells (HLCs) differentiated from pluripotent stem cells by the use of soluble factors can model human liver function and toxicity. However, at present HLC maturity and whether any deficit represents a true fetal state or aberrant differentiation is unclear and compounded by comparison to potentially deteriorated adult hepatocytes. Therefore, we generated HLCs from multiple lineages and two different protocols for direct comparison with fresh fetal and adult hepatocytes. METHODS: Protocols were developed for robust differentiation. Multiple transcript, protein and functional analyses compared HLCs to fresh human fetal and adult hepatocytes. RESULTS: HLCs were comparable to those of other laboratories by multiple parameters. Transcriptional changes during differentiation mimicked human embryogenesis and showed more similarity to pericentral than periportal hepatocytes. Unbiased proteomics demonstrated greater proximity to liver than 30 other human organs or tissues. However, by comparison to fresh material HLC maturity was proven by transcript, protein and function to be fetal-like and short of the adult phenotype. Expression of 81% of phase 1 enzymes in HLCs was significantly upregulated and half were statistically no different from in fetal hepatocytes. HLCs secreted albumin and metabolized testosterone (CYP3A) and dextrorphan (CYP2D6) like fetal hepatocytes. In seven bespoke tests devised by principal components analysis to distinguish fetal from adult hepatocytes HLCs from two different source laboratories consistently demonstrated fetal characteristics. CONCLUSIONS: HLCs from different sources are broadly comparable with unbiased proteomic evidence for faithful differentiation down the liver lineage. This current phenotype mimics human fetal rather than adult hepatocytes.

Original language	English
Pages (from-to)	581-589
Number of pages	8
Journal	Journal of Hepatology
Volume	62
Issue number	3
Early online date	18 Oct 2014
DOIs	https://doi.org/10.1016/j.jhep.2014.10.016
Publication status	Published - Mar 2015

Keywords

Embryo
Hepatic
Hepatotoxicity
Human embryonic stem cell
Liver

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Baxter, M., Withey, S., Harrison, S., Segeritz, C.-P., Zhang, F., Atkinson-Dell, R., Rowe, C., Gerrard, D., Sison-Young, R., Jenkins, R., Henry, J., Berry, A., Mohamet, L., Best, M., Fenwick, S. W., Malik, H., Kitteringham, N. R., Goldring, C. E., Piper Hanley, K., ... Hanley, N. (2015). Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes. Journal of Hepatology, 62(3), 581-589. https://doi.org/10.1016/j.jhep.2014.10.016

@article{6db2094f76384535bb83016db2cec9d4,

title = "Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes",

abstract = "BACKGROUND & AIMS: Hepatocyte-like cells (HLCs) differentiated from pluripotent stem cells by the use of soluble factors can model human liver function and toxicity. However, at present HLC maturity and whether any deficit represents a true fetal state or aberrant differentiation is unclear and compounded by comparison to potentially deteriorated adult hepatocytes. Therefore, we generated HLCs from multiple lineages and two different protocols for direct comparison with fresh fetal and adult hepatocytes. METHODS: Protocols were developed for robust differentiation. Multiple transcript, protein and functional analyses compared HLCs to fresh human fetal and adult hepatocytes. RESULTS: HLCs were comparable to those of other laboratories by multiple parameters. Transcriptional changes during differentiation mimicked human embryogenesis and showed more similarity to pericentral than periportal hepatocytes. Unbiased proteomics demonstrated greater proximity to liver than 30 other human organs or tissues. However, by comparison to fresh material HLC maturity was proven by transcript, protein and function to be fetal-like and short of the adult phenotype. Expression of 81% of phase 1 enzymes in HLCs was significantly upregulated and half were statistically no different from in fetal hepatocytes. HLCs secreted albumin and metabolized testosterone (CYP3A) and dextrorphan (CYP2D6) like fetal hepatocytes. In seven bespoke tests devised by principal components analysis to distinguish fetal from adult hepatocytes HLCs from two different source laboratories consistently demonstrated fetal characteristics. CONCLUSIONS: HLCs from different sources are broadly comparable with unbiased proteomic evidence for faithful differentiation down the liver lineage. This current phenotype mimics human fetal rather than adult hepatocytes.",

keywords = "Embryo, Hepatic, Hepatotoxicity, Human embryonic stem cell, Liver",

author = "Melissa Baxter and Sarah Withey and Sean Harrison and Charis-P Segeritz and Fang Zhang and Rebecca Atkinson-Dell and Cliff Rowe and David Gerrard and Rowena Sison-Young and Roz Jenkins and Joanne Henry and Andrew Berry and Lisa Mohamet and Marie Best and Fenwick, {Stephen W.} and Hassan Malik and Kitteringham, {Neil R.} and Goldring, {Chris E.} and {Piper Hanley}, Karen and Ludovic Vallier and Neil Hanley",

year = "2015",

month = mar,

doi = "10.1016/j.jhep.2014.10.016",

language = "English",

volume = "62",

pages = "581--589",

journal = "Journal of Hepatology",

issn = "0168-8278",

publisher = "Elsevier BV",

number = "3",

}

Baxter, M, Withey, S, Harrison, S, Segeritz, C-P, Zhang, F, Atkinson-Dell, R, Rowe, C, Gerrard, D, Sison-Young, R, Jenkins, R, Henry, J, Berry, A, Mohamet, L, Best, M, Fenwick, SW, Malik, H, Kitteringham, NR, Goldring, CE, Piper Hanley, K, Vallier, L & Hanley, N 2015, 'Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes', Journal of Hepatology, vol. 62, no. 3, pp. 581-589. https://doi.org/10.1016/j.jhep.2014.10.016

TY - JOUR

T1 - Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes

AU - Baxter, Melissa

AU - Withey, Sarah

AU - Harrison, Sean

AU - Segeritz, Charis-P

AU - Zhang, Fang

AU - Atkinson-Dell, Rebecca

AU - Rowe, Cliff

AU - Gerrard, David

AU - Sison-Young, Rowena

AU - Jenkins, Roz

AU - Henry, Joanne

AU - Berry, Andrew

AU - Mohamet, Lisa

AU - Best, Marie

AU - Fenwick, Stephen W.

AU - Malik, Hassan

AU - Kitteringham, Neil R.

AU - Goldring, Chris E.

AU - Piper Hanley, Karen

AU - Vallier, Ludovic

AU - Hanley, Neil

PY - 2015/3

Y1 - 2015/3

N2 - BACKGROUND & AIMS: Hepatocyte-like cells (HLCs) differentiated from pluripotent stem cells by the use of soluble factors can model human liver function and toxicity. However, at present HLC maturity and whether any deficit represents a true fetal state or aberrant differentiation is unclear and compounded by comparison to potentially deteriorated adult hepatocytes. Therefore, we generated HLCs from multiple lineages and two different protocols for direct comparison with fresh fetal and adult hepatocytes. METHODS: Protocols were developed for robust differentiation. Multiple transcript, protein and functional analyses compared HLCs to fresh human fetal and adult hepatocytes. RESULTS: HLCs were comparable to those of other laboratories by multiple parameters. Transcriptional changes during differentiation mimicked human embryogenesis and showed more similarity to pericentral than periportal hepatocytes. Unbiased proteomics demonstrated greater proximity to liver than 30 other human organs or tissues. However, by comparison to fresh material HLC maturity was proven by transcript, protein and function to be fetal-like and short of the adult phenotype. Expression of 81% of phase 1 enzymes in HLCs was significantly upregulated and half were statistically no different from in fetal hepatocytes. HLCs secreted albumin and metabolized testosterone (CYP3A) and dextrorphan (CYP2D6) like fetal hepatocytes. In seven bespoke tests devised by principal components analysis to distinguish fetal from adult hepatocytes HLCs from two different source laboratories consistently demonstrated fetal characteristics. CONCLUSIONS: HLCs from different sources are broadly comparable with unbiased proteomic evidence for faithful differentiation down the liver lineage. This current phenotype mimics human fetal rather than adult hepatocytes.

AB - BACKGROUND & AIMS: Hepatocyte-like cells (HLCs) differentiated from pluripotent stem cells by the use of soluble factors can model human liver function and toxicity. However, at present HLC maturity and whether any deficit represents a true fetal state or aberrant differentiation is unclear and compounded by comparison to potentially deteriorated adult hepatocytes. Therefore, we generated HLCs from multiple lineages and two different protocols for direct comparison with fresh fetal and adult hepatocytes. METHODS: Protocols were developed for robust differentiation. Multiple transcript, protein and functional analyses compared HLCs to fresh human fetal and adult hepatocytes. RESULTS: HLCs were comparable to those of other laboratories by multiple parameters. Transcriptional changes during differentiation mimicked human embryogenesis and showed more similarity to pericentral than periportal hepatocytes. Unbiased proteomics demonstrated greater proximity to liver than 30 other human organs or tissues. However, by comparison to fresh material HLC maturity was proven by transcript, protein and function to be fetal-like and short of the adult phenotype. Expression of 81% of phase 1 enzymes in HLCs was significantly upregulated and half were statistically no different from in fetal hepatocytes. HLCs secreted albumin and metabolized testosterone (CYP3A) and dextrorphan (CYP2D6) like fetal hepatocytes. In seven bespoke tests devised by principal components analysis to distinguish fetal from adult hepatocytes HLCs from two different source laboratories consistently demonstrated fetal characteristics. CONCLUSIONS: HLCs from different sources are broadly comparable with unbiased proteomic evidence for faithful differentiation down the liver lineage. This current phenotype mimics human fetal rather than adult hepatocytes.

KW - Embryo

KW - Hepatic

KW - Hepatotoxicity

KW - Human embryonic stem cell

KW - Liver

U2 - 10.1016/j.jhep.2014.10.016

DO - 10.1016/j.jhep.2014.10.016

M3 - Article

C2 - 25457200

SN - 0168-8278

VL - 62

SP - 581

EP - 589

JO - Journal of Hepatology

JF - Journal of Hepatology

IS - 3

ER -

Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes

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