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Generation of Liver Disease-Specific Induced Pluripotent Stem Cells Along with Efficient Differentiation to Functional Hepatocyte-Like Cells

Stem Cell Reviews and Reports volume 6, pages 622–632 (2010)Cite this article

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Abstract

The availability of disease-specific induced pluripotent stem cells (iPSCs) offers a unique opportunity for studying and modeling the effects of specific gene defects on human liver development in vitro and for testing small molecules or other potential therapies for relevant liver disorders. Here we report, for the first time, the derivation of iPSCs by the retroviral transduction of Yamanaka’s factors in serum and feeder-free culture conditions from liver-specific patients with tyrosinemia, glycogen storage disease, progressive familial hereditary cholestasis, and two siblings with Crigler-Najjar syndrome. Furthermore, they were differentiated into functional hepatocyte-like cells efficiently. These iPSCs possessed properties of human embryonic stem cells (hESCs) and were successfully differentiated into three lineages that resembled hESC morphology, passaging, surface and pluripotency markers, normal karyotype, DNA methylation, and differentiation. The hepatic lineage-directed differentiation showed that the iPSC-derived hepatic cells expressed hepatocyte-specific markers. Their functionality was confirmed by glycogen and lipid storage activity, secretion of albumin, alpha-fetoprotein, and urea, CYP450 metabolic activity, as well as LDL and indocyanin green uptake. Our results provide proof of principal that human liver-disease specific iPSCs present an exciting potential venue toward cell-based therapeutics, drug metabolism, human liver development and disease models for liver failure disorders.

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Change history

Abbreviations

hiPSCs:

Human induced pluripotent stem cells

hESCs:

Human embryonic stem cells

TYR:

Tyrosinemia

GSD:

Glycogen storage disease

HER:

Hereditary cholestasis

CNS:

Crigler-Najjar syndrome

EB:

Embryoid body

AFP:

Alpha-fetoprotein

ALB:

Albumin

LDL:

Low-density lipoprotein

PAS:

Periodic Acid-Schiff

ICG:

Indocyanin green

HDFs:

Human dermal fibroblasts

HLCs:

Hepatocyte like cells

ALP:

Alkaline phosphatase

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Acknowledgements

This study was funded by a grant provided from Royan Institute and Iranian Stem Cell Council.

Financial support

This study was funded by a grant provided from Royan Institute and Iranian Stem Cell Council.

Disclosures

None of the authors have any conflicts of interest to disclose and all authors support submission to this journal.

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Authors and Affiliations

  1. Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, Iran

    Arefeh Ghodsizadeh, Adeleh Taei, Ali Seifinejad, Behshad Pournasr, Nasser Aghdami & Hossein Baharvand

  2. Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran

    Arefeh Ghodsizadeh

  3. Department of Genetics, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Mehdi Totonchi, Hamid Gourabi & Navid Almadani

  4. Department of Regenerative Biomedicine, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Nasser Aghdami

  5. Digestive Disease Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran

    Reza Malekzadeh

  6. Department of Molecular Systems Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Ghasem Hosseini Salekdeh

  7. Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran

    Hossein Baharvand

Authors
  1. Arefeh Ghodsizadeh
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  2. Adeleh Taei
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  3. Mehdi Totonchi
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  4. Ali Seifinejad
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  5. Hamid Gourabi
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  6. Behshad Pournasr
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  7. Nasser Aghdami
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  8. Reza Malekzadeh
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  9. Navid Almadani
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  10. Ghasem Hosseini Salekdeh
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  11. Hossein Baharvand
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Corresponding author

Correspondence to Hossein Baharvand.

Additional information

Arefeh Ghodsizadeh and Adeleh Taei contributed equally in this work.

Electronic supplementary material

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Supplementary Table 1

Relative mRNA levels of lineage-specific hepatic markers [albumin (ALB), hepatic nuclear factor 4α (HNF4α) and CYP3a4], endoderm markers (SOX 17 and AFP), and markers for undifferentiated cells (Oct4) in differentiated and undifferentiated states. Data are normalized to β-actin and depicted as mean ± standard error of mean. (DOC 31 kb)

Supplementary Table 2

Primer sequences and conditions of real time RT-PCR. (DOC 37 kb)

Supplementary Fig. 1

A schematic view of protocol for the directed differentiation of hiPSCs toward HLCs. (DOC 107 kb)

Supplementary Fig. 2

PROD assay on hiPSC-HLCs of three lines at day 18, before and after CYP450 induction with phenobarbital. (DOC 501 kb)

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Ghodsizadeh, A., Taei, A., Totonchi, M. et al. Generation of Liver Disease-Specific Induced Pluripotent Stem Cells Along with Efficient Differentiation to Functional Hepatocyte-Like Cells. Stem Cell Rev and Rep 6, 622–632 (2010). https://doi.org/10.1007/s12015-010-9189-3

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  • DOI: https://doi.org/10.1007/s12015-010-9189-3

Keywords

  • Differentiation
  • Hepatocyte
  • Human induced pluripotent stem cells
  • Liver disease