Stem Cell Reviews and Reports

, Volume 6, Issue 4, pp 622–632 | Cite as

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

  • Arefeh Ghodsizadeh
  • Adeleh Taei
  • Mehdi Totonchi
  • Ali Seifinejad
  • Hamid Gourabi
  • Behshad Pournasr
  • Nasser Aghdami
  • Reza Malekzadeh
  • Navid Almadani
  • Ghasem Hosseini Salekdeh
  • Hossein Baharvand
Article

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.

Keywords

Differentiation Hepatocyte Human induced pluripotent stem cells Liver disease 

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

Notes

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.

Supplementary material

12015_2010_9189_MOESM1_ESM.doc (32 kb)
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)
12015_2010_9189_MOESM2_ESM.doc (38 kb)
Supplementary Table 2 Primer sequences and conditions of real time RT-PCR. (DOC 37 kb)
12015_2010_9189_MOESM3_ESM.doc (108 kb)
Supplementary Fig. 1 A schematic view of protocol for the directed differentiation of hiPSCs toward HLCs. (DOC 107 kb)
12015_2010_9189_MOESM4_ESM.doc (501 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)
ESM 5

(MP4 1746 kb)

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Arefeh Ghodsizadeh
    • 1
    • 2
  • Adeleh Taei
    • 1
  • Mehdi Totonchi
    • 3
  • Ali Seifinejad
    • 1
  • Hamid Gourabi
    • 3
  • Behshad Pournasr
    • 1
  • Nasser Aghdami
    • 1
    • 4
  • Reza Malekzadeh
    • 5
  • Navid Almadani
    • 3
  • Ghasem Hosseini Salekdeh
    • 6
  • Hossein Baharvand
    • 1
    • 7
  1. 1.Department of Stem Cells and Developmental BiologyRoyan Institute for Stem Cell Biology and Technology, ACECRTehranIran
  2. 2.Department of Biotechnology, College of ScienceUniversity of TehranTehranIran
  3. 3.Department of GeneticsRoyan Institute for Reproductive Biomedicine, ACECRTehranIran
  4. 4.Department of Regenerative BiomedicineRoyan Institute for Stem Cell Biology and Technology, ACECRTehranIran
  5. 5.Digestive Disease Research Center, Shariati HospitalTehran University of Medical SciencesTehranIran
  6. 6.Department of Molecular Systems BiologyRoyan Institute for Stem Cell Biology and Technology, ACECRTehranIran
  7. 7.Department of Developmental BiologyUniversity of Science and Culture, ACECRTehranIran

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