Generation of Liver Disease-Specific Induced Pluripotent Stem Cells Along with Efficient Differentiation to Functional Hepatocyte-Like Cells
First Online: 07 September 2010 DOI:
Cite this article as: Ghodsizadeh, A., Taei, A., Totonchi, M. et al. Stem Cell Rev and Rep (2010) 6: 622. doi:10.1007/s12015-010-9189-3 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
Human embryonic stem cells
Glycogen storage disease
Human dermal fibroblasts
Hepatocyte like cells
Arefeh Ghodsizadeh and Adeleh Taei contributed equally in this work.
Electronic supplementary material
The online version of this article (doi:
) contains supplementary material, which is available to authorized users. 10.1007/s12015-010-9189-3 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_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) References
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