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Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix

Nature Medicine volume 16, pages 814–820 (2010)Cite this article

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Abstract

Orthotopic liver transplantation is the only available treatment for severe liver failure, but it is currently limited by organ shortage. One technical challenge that has thus far limited the development of a tissue-engineered liver graft is oxygen and nutrient transport. Here we demonstrate a novel approach to generate transplantable liver grafts using decellularized liver matrix. The decellularization process preserves the structural and functional characteristics of the native microvascular network, allowing efficient recellularization of the liver matrix with adult hepatocytes and subsequent perfusion for in vitro culture. The recellularized graft supports liver-specific function including albumin secretion, urea synthesis and cytochrome P450 expression at comparable levels to normal liver in vitro. The recellularized liver grafts can be transplanted into rats, supporting hepatocyte survival and function with minimal ischemic damage. These results provide a proof of principle for the generation of a transplantable liver graft as a potential treatment for liver disease.

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Figure 1: Decellularization of ischemic rat livers.
Figure 2: DLM retains intact lobular structure and vascular bed.
Figure 3: Repopulation of rat DLM with adult rat hepatocytes.
Figure 4: Hepatic function of the recellularized liver graft in vitro.
Figure 5: Transplantation of the recellularized liver graft.

References

  1. Heron, M. et al. Deaths: Final Data for 2006. in National Vital Statistics Report, Vol. 57 (Centers for Disease Control and Prevention, Hyattsville, Maryland, 2009).

  2. Punch, J.D., Hayes, D.H., LaPorte, F.B., McBride, V. & Seely, M.S. Organ donation and utilization in the United States, 1996–2005. Am. J. Transplant. 7, 1327–1338 (2007).

    Article  CAS  Google Scholar 

  3. Fox, I.J. & Roy-Chowdhury, J. Hepatocyte transplantation. J. Hepatol. 40, 878–886 (2004).

    Article  CAS  Google Scholar 

  4. Strom, S.C. et al. Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure. Transplantation 63, 559–569 (1997).

    Article  CAS  Google Scholar 

  5. Horslen, S.P. et al. Isolated hepatocyte transplantation in an infant with a severe urea cycle disorder. Pediatrics 111, 1262–1267 (2003).

    Article  Google Scholar 

  6. Fox, I.J. et al. Treatment of the Crigler-Najjar syndrome type I with hepatocyte transplantation. N. Engl. J. Med. 338, 1422–1426 (1998).

    Article  CAS  Google Scholar 

  7. Fisher, R.A. & Strom, S.C. Human hepatocyte transplantation: worldwide results. Transplantation 82, 441–449 (2006).

    Article  Google Scholar 

  8. Habibullah, C.M., Syed, I.H., Qamar, A. & Taher-Uz, Z. Human fetal hepatocyte transplantation in patients with fulminant hepatic failure. Transplantation 58, 951–952 (1994).

    Article  CAS  Google Scholar 

  9. Nagata, H. et al. Prolonged survival of porcine hepatocytes in cynomolgus monkeys. Gastroenterology 132, 321–329 (2007).

    Article  CAS  Google Scholar 

  10. Basma, H. et al. Differentiation and transplantation of human embryonic stem cell–derived hepatocytes. Gastroenterology 136, 990–999 (2009).

    Article  CAS  Google Scholar 

  11. Cho, C.H. et al. Homogeneous differentiation of hepatocyte-like cells from embryonic stem cells: applications for the treatment of liver failure. FASEB J. 22, 898–909 (2008).

    Article  CAS  Google Scholar 

  12. Totsugawa, T. et al. Survival of liver failure pigs by transplantation of reversibly immortalized human hepatocytes with tamoxifen-mediated self-recombination. J. Hepatol. 47, 74–82 (2007).

    Article  CAS  Google Scholar 

  13. Shafritz, D.A., Oertel, M., Menthena, A., Nierhoff, D. & Dabeva, M.D. Liver stem cells and prospects for liver reconstitution by transplanted cells. Hepatology 43, S89–S98 (2006).

    Article  CAS  Google Scholar 

  14. Smets, F., Najimi, M. & Sokal, E.M. Cell transplantation in the treatment of liver diseases. Pediatr. Transplant. 12, 6–13 (2008).

    Article  CAS  Google Scholar 

  15. Langer, R. & Vacanti, J.P. Tissue engineering. Science 260, 920–926 (1993).

    Article  CAS  Google Scholar 

  16. Wang, S., Nagrath, D., Chen, P.C., Berthiaume, F. & Yarmush, M.L. Three-dimensional primary hepatocyte culture in synthetic self-assembling peptide hydrogel. Tissue. Eng. Part A 14, 227–236 (2008).

    Article  CAS  Google Scholar 

  17. Nahmias, Y., Berthiaume, F. & Yarmush, M.L. Integration of technologies for hepatic tissue engineering. Adv. Biochem. Eng. Biotechnol. 103, 309–329 (2007).

    PubMed  Google Scholar 

  18. Ohashi, K. et al. Engineering functional two- and three-dimensional liver systems in vivo using hepatic tissue sheets. Nat. Med. 13, 880–885 (2007).

    Article  CAS  Google Scholar 

  19. Kulig, K.M. & Vacanti, J.P. Hepatic tissue engineering. Transpl. Immunol. 12, 303–310 (2004).

    Article  CAS  Google Scholar 

  20. Nahmias, Y., Schwartz, R.E., Hu, W.S., Verfaillie, C.M. & Odde, D.J. Endothelium-mediated hepatocyte recruitment in the establishment of liver-like tissue in vitro. Tissue Eng. 12, 1627–1638 (2006).

    Article  CAS  Google Scholar 

  21. Badylak, S.F. The extracellular matrix as a biologic scaffold material. Biomaterials 28, 3587–3593 (2007).

    Article  CAS  Google Scholar 

  22. Yoo, J.J., Meng, J., Oberpenning, F. & Atala, A. Bladder augmentation using allogenic bladder submucosa seeded with cells. Urology 51, 221–225 (1998).

    Article  CAS  Google Scholar 

  23. Dahl, S.L., Koh, J., Prabhakar, V. & Niklason, L.E. Decellularized native and engineered arterial scaffolds for transplantation. Cell Transplant. 12, 659–666 (2003).

    Article  Google Scholar 

  24. Nieponice, A., Gilbert, T.W. & Badylak, S.F. Reinforcement of esophageal anastomoses with an extracellular matrix scaffold in a canine model. Ann. Thorac. Surg. 82, 2050–2058 (2006).

    Article  Google Scholar 

  25. Schechner, J.S. et al. Engraftment of a vascularized human skin equivalent. FASEB J. 17, 2250–2256 (2003).

    Article  CAS  Google Scholar 

  26. Macchiarini, P. et al. Clinical transplantation of a tissue-engineered airway. Lancet 372, 2023–2030 (2008).

    Article  Google Scholar 

  27. Ott, H.C. et al. Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart. Nat. Med. 14, 213–221 (2008).

    Article  CAS  Google Scholar 

  28. Safar, P. Clinical death symposium. Crit. Care Med. 16, 919–920 (1988).

    Article  CAS  Google Scholar 

  29. Hoffenberg, R. Measurement of the synthesis of liver-produced plasma proteins with particular reference to dietary protein and amino acid supply. Biochem. J. 129, 3P (1972).

    Article  CAS  Google Scholar 

  30. Khetani, S.R. & Bhatia, S.N. Microscale culture of human liver cells for drug development. Nat. Biotechnol. 26, 120–126 (2008).

    Article  CAS  Google Scholar 

  31. Tolboom, H. et al. Recovery of warm ischemic rat liver grafts by normothermic extracorporeal perfusion. Transplantation 87, 170–177 (2009).

    Article  Google Scholar 

  32. Uygun, K. et al. Diluted blood reperfusion as a model for transplantation of ischemic rat livers: alt is a direct indicator of viability. Transplant. Proc. (in the press).

  33. Cho, C.H. et al. Oxygen uptake rates and liver-specific functions of hepatocyte and 3T3 fibroblast co-cultures. Biotechnol. Bioeng. 97, 188–199 (2007).

    Article  CAS  Google Scholar 

  34. Rotem, A. et al. Oxygen is a factor determining in vitro tissue assembly: effects on attachment and spreading of hepatocytes. Biotechnol. Bioeng. 43, 654–660 (1994).

    Article  CAS  Google Scholar 

  35. Foy, B.D. et al. Optimization of hepatocyte attachment to microcarriers: importance of oxygen. Biotechnol. Bioeng. 42, 579–588 (1993).

    Article  CAS  Google Scholar 

  36. Balis, U.J. et al. Oxygen consumption characteristics of porcine hepatocytes. Metab. Eng. 1, 49–62 (1999).

    Article  CAS  Google Scholar 

  37. Dunn, J.C., Tompkins, R.G. & Yarmush, M.L. Long-term in vitro function of adult hepatocytes in a collagen sandwich configuration. Biotechnol. Prog. 7, 237–245 (1991).

    Article  CAS  Google Scholar 

  38. Schleimer, K. et al. Improved technique of heterotopic auxiliary rat liver transplantation with portal vein arterialization. Langenbecks Arch. Surg. 391, 102–107 (2006).

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the US National Institutes of Health, R01DK59766 and R01DK084053 to M.L.Y., K99/R00 DK080942 to K.U. and K99DK083556 to A.S.-G., and US National Science Foundation CBET-0853569 to K.U. We thank the support of the Shriners Hospitals for Children to B.E.U. (grant no. 8503), the American Liver Foundation to A.S.-G. and Massachusetts General Hospital Junior Faculty Grant to K.U. and the Shriners Hospitals for Children. We would like to thank A. Vitalo, C. Calhoun and B. Crowther for technical support.

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Author notes

  1. Alejandro Soto-Gutierrez, Hiroshi Yagi & Yaakov Nahmias

    Present address: Current addresses: Center for Innovative Regenerative Therapies, Department of Surgery, Transplantation Section, Children's Hospital of Pittsburgh, McGowan Institute for Regenerative Medicine and University of Pittsburgh, Pittsburgh, Pennsylvania, USA (A.S.-G.), Department of Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan (H.Y.) and the Selim and Rachel Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel (Y.N.).,

Authors and Affiliations

  1. Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, Massachusetts, USA

    Basak E Uygun, Alejandro Soto-Gutierrez, Hiroshi Yagi, Maria-Louisa Izamis, Maria A Guzzardi, Carley Shulman, Jack Milwid, Arno Tilles, Francois Berthiaume, Yaakov Nahmias, Martin L Yarmush & Korkut Uygun

  2. Sector of Medicine, Scuola Superiore Sant'Anna, Pisa, Italy

    Maria A Guzzardi

  3. Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Shikata-cho, Okayama, Japan.,

    Naoya Kobayashi

  4. Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA

    Francois Berthiaume & Martin L Yarmush

  5. Transplantation Unit, Massachusetts General Hospital, Boston, Massachusetts, USA

    Martin Hertl

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  1. Basak E Uygun
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Contributions

K.U. had full access to all of the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analyses. B.E.U., A.S.-G., H.Y. and K.U., study concept and design; B.E.U., A.S.-G., H.Y., M.A.G., acquisition of data; B.E.U., A.S.-G., Y.N. and K.U., analysis and interpretation of data; M.-L.I., C.S. and B.E.U., rat liver harvest, decellularization and hepatocyte isolation; J.M. and B.E.U., design and construction of the recellularized liver chamber; B.E.U., A.S.-G., H.Y. and M.A.G., recellularization; A.S.-G. and H.Y., histology and transplantation studies; B.E.U., A.S.-G., H.Y. and K.U., drafting of the manuscript; B.E.U., A.S.-G., H.Y., M.L.Y., Y.N., A.T., F.B., M.H., N.K. and K.U., critical revision of the manuscript for intellectual content; B.E.U. and K.U., statistical analysis; M.L.Y., K.U. and A.S.-G., obtained funding; M.-L.I., C.S., J.M., Y.N., A.T. and F.B., administrative, technical or material support. All authors contributed to the preparation of the report.

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Correspondence to Korkut Uygun.

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Supplementary Tables 1 and 2, Supplementary Figures 1–5 and Supplementary Methods (PDF 4275 kb)

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Uygun, B., Soto-Gutierrez, A., Yagi, H. et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med 16, 814–820 (2010). https://doi.org/10.1038/nm.2170

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