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Vascularized and functional human liver from an iPSC-derived organ bud transplant

Nature volume 499, pages 481–484 (2013)Cite this article

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Abstract

A critical shortage of donor organs for treating end-stage organ failure highlights the urgent need for generating organs from human induced pluripotent stem cells (iPSCs)1. Despite many reports describing functional cell differentiation2,3,4, no studies have succeeded in generating a three-dimensional vascularized organ such as liver. Here we show the generation of vascularized and functional human liver from human iPSCs by transplantation of liver buds created in vitro (iPSC-LBs). Specified hepatic cells (immature endodermal cells destined to track the hepatic cell fate) self-organized into three-dimensional iPSC-LBs by recapitulating organogenetic interactions between endothelial and mesenchymal cells5. Immunostaining and gene-expression analyses revealed a resemblance between in vitro grown iPSC-LBs and in vivo liver buds. Human vasculatures in iPSC-LB transplants became functional by connecting to the host vessels within 48 hours. The formation of functional vasculatures stimulated the maturation of iPSC-LBs into tissue resembling the adult liver. Highly metabolic iPSC-derived tissue performed liver-specific functions such as protein production and human-specific drug metabolism without recipient liver replacement6. Furthermore, mesenteric transplantation of iPSC-LBs rescued the drug-induced lethal liver failure model. To our knowledge, this is the first report demonstrating the generation of a functional human organ from pluripotent stem cells. Although efforts must ensue to translate these techniques to treatments for patients, this proof-of-concept demonstration of organ-bud transplantation provides a promising new approach to study regenerative medicine.

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Figure 1: Generation of human liver buds from human iPSCs.
Figure 2: In vitro characterization of human iPSC-LBs.
Figure 3: Generation of human liver with functional vascular networks in vivo.
Figure 4: Functional characterization of human iPSC-LB derived liver.

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Gene Expression Omnibus

Data deposits

Microarray data, including that of human iPSC-LBs, human FLC-LBs, human adult liver tissues (ALT) and mouse liver tissue of various developmental stages, have been deposited in the Gene Expression Omnibus under accession number GSE46631.

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Acknowledgements

We thank F. Kawamata, E. Yoshizawa, Y. Suzuki, S. Nakai, Y. Takahashi, N. Tsuchida and N. Sasaki for kindly providing technical support; J. Nakabayashi, K. Yasumura, R. Fujiwara, T. Amiya, A. Nakano, Y. Mitsuhashi and all of the members of our laboratory for help with several experiments and comments. We are also grateful to D. Fukumura, Y. Goshima, T. Hirose, M. Ichino, U. Yokoyama, T. Ogawa and R. K. Jain for critical evaluation of the manuscript. This work was supported by grants to H. Taniguchi from the Strategic Promotion of Innovative Research and Development (S-innovation, 62890004) of the Japan Science and Technology Agency (JST). This work was also supported by the Grants-in-Aid of the Ministry of Education, Culture, Sports, Science, and Technology of Japan to T. Takebe (no. 24106510, 24689052), N. Koike (no. 22390260) and H. Taniguchi (no. 21249071, 25253079); by the Specified Research Grant of the Takeda Science Foundation and a grant from the Japan IDDM network to H. Taniguchi; and by a grant of the Yokohama Foundation for Advanced Medical Science to T. Takebe.

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Authors and Affiliations

  1. Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan,

    Takanori Takebe, Keisuke Sekine, Masahiro Enomura, Hiroyuki Koike, Masaki Kimura, Takunori Ogaeri, Ran-Ran Zhang, Yasuharu Ueno, Yun-Wen Zheng, Naoto Koike & Hideki Taniguchi

  2. Advanced Medical Research Center, Yokohama City University, Yokohama, Kanagawa 236-0004, Japan,

    Takanori Takebe & Hideki Taniguchi

  3. Department of Surgery, Seirei Sakura Citizen Hospital, 2-36-2 Ebaradai, Sakura, Chiba 285-8765, Japan,

    Naoto Koike

  4. ADME & Tox. Research Institute, Sekisui Medical Company Ltd., Tokai, Ibaraki 319-1182, Japan,

    Shinsuke Aoyama & Yasuhisa Adachi

Authors
  1. Takanori Takebe
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Contributions

T.T. conceived the study, performed the experiments, collected and analysed the data and drafted the manuscript. K.S., M.E., H.K., M.K., T.O., R-R.Z. and S.A. performed the experiments with the technical guidance and expertise of K.S., Y.-W.Z., Y.U. and T.T. K.S., Y.-W.Z., N.K., Y.A. and H.T. provided critical advice on the research strategy and design.

Corresponding authors

Correspondence to Takanori Takebe or Hideki Taniguchi.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-20, Supplementary Methods, Supplementary Tables 1-3, Supplementary Discussion and Supplementary References. This file was replaced on 27 November 2013 to include the Supplementary Discussion and Tables. (PDF 9046 kb)

Generation of human induced pluripotent stem cell-derived liver bud by recapitulating organogenesis

This video shows formation of human induced pluripotent stem cell-derived liver bud by recapitulating organogenetic interactions. (MOV 7371 kb)

Formation of functional human vascular networks inside the hiPSC-LB transplant

This video shows the z stack images of patent human vasculatures inside the hiPSC-LB transplants at day 4. (MOV 3646 kb)

Successful engraftment of hiPSC-derived hepatic cells through the functional vessel formation

This video shows the successful engraftment of hiPSC-Heps along with the functional vascular networks at day 4. (MOV 3968 kb)

3D visualization of the direct connection among human and host vessels by whole mount immunostaining

This video shows the connection among the human and host vessels visualised by immunostaining of the explants. Volocity software was used to reconstruct 3D image. (MOV 5222 kb)

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Takebe, T., Sekine, K., Enomura, M. et al. Vascularized and functional human liver from an iPSC-derived organ bud transplant. Nature 499, 481–484 (2013). https://doi.org/10.1038/nature12271

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