Generation and Hepatic Differentiation of Human iPS Cells

  • Tetsuya IshikawaEmail author
  • Keitaro Hagiwara
  • Takahiro Ochiya
Part of the Methods in Molecular Biology book series (MIMB, volume 826)


A method for the generation of human induced pluripotent stem (iPS) cells was established. This method employs adenovirus carrying the ecotropic retrovirus receptor mCAT1 and Moloney murine leukemia virus (MMLV)-based retroviral vectors carrying the four transcription factors POU5F1 (OCT3/4), KLF4, SOX2, and MYC (c-Myc) (Masaki H & Ishikawa T Stem Cell Res 1:105–15, 2007). The differentiation of human iPS cells into hepatic cells was performed by a stepwise protocol (Song Z et al. Cell Res 19:1233–42, 2009). These cells have potential as patient-specific in vitro models for studying disease etiology and could be used in drug discovery programs tailored to deal with genetic variations in drug efficacy and toxicity.

Key words

Human iPS cells POU5F1 (OCT3/4) KLF4 Sox2 MYC (c-Myc) Retrovirus Hepatic differentiation 



This work was supported in part by a Grant-in-Aid for the Third-Term Comprehensive 10-Year Strategy for Cancer Control (T.O.); Health Science Research Grants for Research on the Human Genome and Regenerative Medicine from the Ministry of Health, Labor, and Welfare of Japan (T.O.); and a Grant from the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (NiBio) (T.I.).


  1. 1.
    Masaki H, Ishikawa T, Takahashi S, Okumura M, Sakai N, Haga M, Kominami K, Migita H, McDonald F, Shimada F, Sakurada K. (2007) Heterogeneity of pluripotent marker gene expression in colonies generated in human iPS cell induction culture. Stem Cell Res. 1, 105–115.PubMedCrossRefGoogle Scholar
  2. 2.
    Song Z, Cai J, Liu Y, Zhao D, Yong J, Duo S, Song X, Guo Y, Zhao Y, Qin H, Yin X, Wu C, Che J, Lu S, Ding M, Deng H. (2009) Efficient generation of hepatocyte-like cells from human induced pluripotent stem cells. Cell Res. 19, 1233–1242.PubMedCrossRefGoogle Scholar
  3. 3.
    Kitamura T., Koshino Y., Shibata F., Oki T., Nakajima H., Nosaka T., Kumagai H. (2003) Retrovirus-mediated gene transfer and expression cloning: Powerful tools in functional genomics. Exp. Hematol. 31, 1007–1014.PubMedGoogle Scholar
  4. 4.
    Dvorak P, Emanuelsson K, Fleck RA, Ford A, Gertow K, Gertsenstein M, Gokhale PJ, Hamilton RS, Hampl A, Healy LE, Hovatta O, Hyllner J, Imreh MP, Itskovitz-Eldor J, Jackson J, Johnson JL, Jones M, Kee K, King BL, Knowles BB, Lako M, Lebrin F, Mallon BS, Manning D, Mayshar Y, McKay RD, Michalska AE, Mikkola M, Mileikovsky M, Minger SL, Moore HD, Mummery CL, Nagy A, Nakatsuji N, O’Brien CM, Oh SK, Olsson C, Otonkoski T, Park KY, Passier R, Patel H, Patel M, Pedersen R, Pera MF, Piekarczyk MS, Pera RA, Reubinoff BE, Robins AJ, Rossant J, Rugg-Gunn P, Schulz TC, Semb H, Sherrer ES, Siemen H, Stacey GN, Stojkovic M, Suemori H, Szatkiewicz J, Turetsky T, Tuuri T, van den Brink S, Vintersten K, Vuoristo S, Ward D, Weaver TA, Young LA, Zhang W. (2007) Characterization of human embryonic stem cell lines by the International Stem Cell Initiative. Nat Biotechnol. 25, 803–816.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Tetsuya Ishikawa
    • 1
    Email author
  • Keitaro Hagiwara
    • 2
  • Takahiro Ochiya
    • 2
  1. 1.Core Facilities for Research and Innovative MedicineNational cancer Center Research InstituteTokyoJapan
  2. 2.Division of Molecular and Cellular MedicineNational Cancer Center Research InstituteTokyoJapan

Personalised recommendations