Skip to main content

Advertisement

SpringerLink
Log in

Liver Bioengineering: Promise, Pitfalls, and Hurdles to Overcome

  • Cellular Transplants (G Orlando, Section Editor)
  • Published:
Current Transplantation Reports Aims and scope Submit manuscript

Abstract

Purpose of Review

In this review, we discuss the recent advancements in liver bioengineering and cell therapy and future advancements to improve the field towards clinical applications.

Recent Findings

3D printing, hydrogel-based tissue fabrication, and the use of native decellularized liver extracellular matrix as a scaffold are used to develop whole or partial liver substitutes. The current focus is on developing a functional liver graft through achieving a non-leaky endothelium and a fully constructed bile duct. Use of cell therapy as a treatment is less invasive and less costly compared to transplantation, however, lack of readily available cell sources with low or no immunogenicity and contradicting outcomes of clinical trials are yet to be overcome.

Summary

Liver bioengineering is advancing rapidly through the development of in vitro and in vivo tissue and organ models. Although there are major challenges to overcome, through optimization of the current methods and successful integration of induced pluripotent stem cells, the development of readily available, patient-specific liver substitutes can be achieved.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

Papers of particular interest, published recently, have been highlighted as: • Of importance •• Of major importance

  1. Everhart JE, Ruhl CE. Burden of digestive diseases in the United States part III: liver, biliary tract, and pancreas. Gastroenterology. 2009;136:1134–44.

    Article  PubMed  Google Scholar 

  2. Mokdad AA, Lopez AD, Shahraz S, Lozano R, Mokdad AH, Stanaway J, et al. Liver cirrhosis mortality in 187 countries between 1980 and 2010: a systematic analysis. BMC Med. 2014;12(145).

  3. Ware BR, Khetani SR. Engineered liver platforms for different phases of drug development. Trends Biotechnol. 2017;35:172–83.

    Article  CAS  PubMed  Google Scholar 

  4. Du Y, et al. Mimicking liver sinusoidal structures and functions using a 3D-configured microfluidic chip. Lab Chip. 2017;17:782–94.

    Article  CAS  PubMed  Google Scholar 

  5. Mi S, Yi X, Du Z, Xu Y, Sun W. Construction of a liver sinusoid based on the laminar flow on chip and self-assembly of endothelial cells. Biofabrication. 2018;10:025010.

    Article  CAS  PubMed  Google Scholar 

  6. Wu Q, et al. Establishment of an ex vivo model of nonalcoholic fatty liver disease using a tissue-engineered liver. ACS Biomater Sci Eng. 2018;4:3016–26.

    Article  CAS  Google Scholar 

  7. Griffith LG, Wells A, Stolz DB. Engineering Liver. Hepatol Baltim Md. 2014;60:1426–34.

    Article  Google Scholar 

  8. Mazza G, Al-Akkad W, Rombouts K, Pinzani M. Liver tissue engineering: from implantable tissue to whole organ engineering. Hepatol Commun. 2017;2:131–41.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Atala A, Bauer SB, Soker S, Yoo JJ, Retik AB. Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet. 2006;367:1241–6.

    Article  Google Scholar 

  10. Gonfiotti A, Jaus MO, Barale D, Baiguera S, Comin C, Lavorini F, et al. The first tissue-engineered airway transplantation: 5-year follow-up results. Lancet Lond Engl. 2014;383:238–44.

  11. Shafiee A, Atala A. Tissue engineering: toward a new era of medicine. Annu Rev Med. 2017;68:29–40.

  12. Mohanty S, Sanger K, Heiskanen A, Trifol J, Szabo P, Dufva M, et al. Fabrication of scalable tissue engineering scaffolds with dual-pore microarchitecture by combining 3D printing and particle leaching. Mater Sci Eng C. 2016;61:180–9.

  13. Lee JW, Choi YJ, Yong WJ, Pati F, Shim JH, Kang KS, et al. Development of a 3D cell printed construct considering angiogenesis for liver tissue engineering. Biofabrication. 2016;8:015007.

  14. Kang K, Kim Y, Jeon H, Lee SB, Kim JS, Park SA, et al. Three-dimensional bioprinting of hepatic structures with directly converted hepatocyte-like cells. Tissue Eng Part A. 2018;24:576–83.

  15. Lee H, Han W, Kim H, Ha DH, Jang J, Kim BS, et al. Development of liver decellularized extracellular matrix bioink for three-dimensional cell printing-based liver tissue engineering. Biomacromolecules. 2017;18:1229–37.

  16. Stevens KR, Scull MA, Ramanan V, Fortin CL, Chaturvedi RR, Knouse KA, et al. In situ expansion of engineered human liver tissue in a mouse model of chronic liver disease. Sci Transl Med. 2017;9:eaah5505.

  17. Lewis PL, Su J, Yan M, Meng F, Glaser SS, Alpini GD, et al. Complex bile duct network formation within liver decellularized extracellular matrix hydrogels. Sci Rep. 2018;8:12220.

  18. Ott HC, Matthiesen TS, Goh SK, Black LD, Kren SM, Netoff TI, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med. 2008;14:213–21.

  19. Uygun BE, Soto-Gutierrez A, Yagi H, Izamis ML, Guzzardi MA, Shulman C, et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med. 2010;16:814–20.

  20. Barakat O, Abbasi S, Rodriguez G, Rios J, Wood RP, Ozaki C, et al. Use of decellularized porcine liver for engineering humanized liver organ. J Surg Res. 2012;173:e11–25.

  21. Lang R, Stern MM, Smith L, Liu Y, Bharadwaj S, Liu G, et al. Three-dimensional culture of hepatocytes on porcine liver tissue-derived extracellular matrix. Biomaterials. 2011;32:7042–52.

  22. Kajbafzadeh A-M, Javan-Farazmand N, Monajemzadeh M, Baghayee A. Determining the optimal decellularization and sterilization protocol for preparing a tissue scaffold of a human-sized liver tissue. Tissue Eng Part C Methods. 2013;19:642–51.

    Article  CAS  PubMed  Google Scholar 

  23. Mazza G, Rombouts K, Rennie Hall A, Urbani L, Vinh Luong T, al-Akkad W, et al. Decellularized human liver as a natural 3D-scaffold for liver bioengineering and transplantation. Sci Rep. 2015;5:13079.

  24. Robertson MJ, Soibam B, O’Leary JG, Sampaio LC, Taylor DA. Recellularization of rat liver: an in vitro model for assessing human drug metabolism and liver biology. PLoS One. 2018;13:e0191892.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Ogiso S, Yasuchika K, Fukumitsu K, Ishii T, Kojima H, Miyauchi Y, et al. Efficient recellularisation of decellularised whole-liver grafts using biliary tree and foetal hepatocytes. Sci Rep. 2016;6.

  26. Kojima H, Yasuchika K, Fukumitsu K, Ishii T, Ogiso S, Miyauchi Y, et al. Establishment of practical recellularized liver graft for blood perfusion using primary rat hepatocytes and liver sinusoidal endothelial cells. Am J Transplant. 2018;18:1351–9.

  27. • Devalliere J, Chen Y, Dooley K, Yarmush ML, Uygun BE. Improving functional re-endothelialization of acellular liver scaffold using REDV cell-binding domain. Acta Biomater. 2018;78:151–64 This study shows a novel method of functionalizing the decellularized liver grafts for higher endothelizalization efficiency, rendering such scaffolds applicable for transplantation.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. •• A Mao S. Sustained in vivo perfusion of a re-endothelialized tissue engineered porcine liver. Int J Transplant Res Med. 2017;3 This study shows 72 hour-long in vivo perfusion of decellularized porcine livers after endothelialization, in the absence of anticoagulants.

  29. Yang W, Chen Q, Xia R, Zhang Y, Shuai L, Lai J, et al. A novel bioscaffold with naturally-occurring extracellular matrix promotes hepatocyte survival and vessel patency in mouse models of heterologous transplantation. Biomaterials. 2018;177:52–66.

  30. Hussein KH, Park K-M, Kang K-S, Woo H-M. Biocompatibility evaluation of tissue-engineered decellularized scaffolds for biomedical application. Mater Sci Eng C. 2016;67:766–78.

    Article  CAS  Google Scholar 

  31. Wang Y, et al. Genipin crosslinking reduced the immunogenicity of xenogeneic decellularized porcine whole-liver matrices through regulation of immune cell proliferation and polarization. Sci Rep. 2016;6(24779).

  32. Verstegen MMA, Willemse J, van den Hoek S, Kremers GJ, Luider TM, van Huizen NA, et al. Decellularization of whole human liver grafts using controlled perfusion for transplantable organ bioscaffolds. Stem Cells Dev. 2017;26:1304–15.

  33. Hannan NRF, Segeritz C-P, Touboul T, Vallier L. Production of hepatocyte-like cells from human pluripotent stem cells. Nat Protoc. 2013;8:430–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Siller R, Greenhough S, Naumovska E, Sullivan GJ. Small-molecule-driven hepatocyte differentiation of human pluripotent stem cells. Stem Cell Rep. 2015;4:939–52.

    Article  CAS  Google Scholar 

  35. Kanninen LK, Harjumäki R, Peltoniemi P, Bogacheva MS, Salmi T, Porola P, et al. Laminin-511 and laminin-521-based matrices for efficient hepatic specification of human pluripotent stem cells. Biomaterials. 2016;103:86–100.

  36. Park K-M, Hussein KH, Hong SH, Ahn C, Yang SR, Park SM, et al. Decellularized liver extracellular matrix as promising tools for transplantable bioengineered liver promotes hepatic lineage commitments of induced pluripotent stem cells. Tissue Eng Part A. 2016;22:449–60.

  37. Jaramillo M, Yeh H, Yarmush ML, Uygun BE. Decellularized human liver extracellular matrix (hDLM)-mediated hepatic differentiation of human induced pluripotent stem cells (hIPSCs). J Tissue Eng Regen Med. 2018;12:e1962–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Palakkan AA, Nanda J, Ross JA. Pluripotent stem cells to hepatocytes, the journey so far. Biomed Rep. 2017;6:367–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Mattei G, Magliaro C, Pirone A, Ahluwalia A. Decellularized human liver is too heterogeneous for designing a generic extracellular matrix mimic hepatic scaffold. Artif Organs. 2017;41:E347–55.

    Article  CAS  PubMed  Google Scholar 

  40. Tolosa L, Pareja E, Gómez-Lechón MJ. Clinical application of pluripotent stem cells: an alternative cell-based therapy for treating liver diseases? Transplantation. 2016;100:2548–57.

    Article  CAS  PubMed  Google Scholar 

  41. Fox I. Hepatocyte transplantation. J Hepatol. 2004;40:878–86.

    Article  CAS  PubMed  Google Scholar 

  42. Strom SC, Fisher RA, Thompson MT, Sanyal AJ, Cole PE, Ham JM, et al. Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure. Transplantation. 1997;63:559–69.

  43. Fisher RA, Strom SC. Human hepatocyte transplantation: worldwide results. Transplantation. 2006;82:441–9.

    Article  PubMed  Google Scholar 

  44. Puppi J, Strom SC, Hughes RD, Bansal S, Castell JV, Dagher I, et al. Improving the techniques for human hepatocyte transplantation: report from a consensus meeting in London. Cell Transplant. 2012;21:1–10.

  45. Horner R, Kluge M, Gassner J, Nösser M, Major RD, Reutzel-Selke A, et al. Hepatocyte isolation after laparoscopic liver resection. Tissue Eng Part C Methods. 2016;22:839–46.

  46. •• Uygun BE, Izamis ML, Jaramillo M, Chen Y, Price G, Ozer S, et al. Discarded livers find a new life: engineered liver grafts using hepatocytes recovered from marginal livers. Artif Organs. 2017;41:579–85 This study describes a method of increasing the pool of available human hepatocytes, the most thoroughly tested cells for liver function replacement.

  47. Jitraruch S, Dhawan A, Hughes RD, Filippi C, Lehec SC, Glover L, et al. Cryopreservation of hepatocyte microbeads for clinical transplantation. Cell Transplant. 2017;26:1341–54.

  48. Gramignoli R, Vosough M, Kannisto K, Srinivasan RC, Strom SC. Clinical hepatocyte transplantation: practical limits and possible solutions. Eur Surg Res Eur Chir Forsch Rech Chir Eur. 2015;54:162–77.

    CAS  Google Scholar 

  49. Terry C, Dhawan A, Mitry RR, Lehec SC, Hughes RD. Optimization of the cryopreservation and thawing protocol for human hepatocytes for use in cell transplantation. Liver Transplant Off Publ Am Assoc Study Liver Dis Int Liver Transplant Soc. 2010;16:229–37.

    Google Scholar 

  50. Tolosa L, Bonora-Centelles A, Donato MT, Mirabet V, Pareja E, Negro A, et al. Influence of platelet lysate on the recovery and metabolic performance of cryopreserved human hepatocytes upon thawing. Transplantation. 2011;91:1340–6.

  51. Yoshizumi Y, Yukawa H, Iwaki R, Fujinaka S, Kanou A, Kanou Y, et al. Immunomodulatory effects of adipose tissue-derived stem cells on Concanavalin A-induced acute liver injury in mice. Cell Med. 2017;9:21–33.

  52. Najimi M, Berardis S, el-Kehdy H, Rosseels V, Evraerts J, Lombard C, et al. Human liver mesenchymal stem/progenitor cells inhibit hepatic stellate cell activation: in vitro and in vivo evaluation. Stem Cell Res Ther. 2017;8:131.

  53. •• Zhao T, Zhang Z-N, Rong Z, Xu Y. Immunogenicity of induced pluripotent stem cells. Nature. 2011;474:212–5 It was shown that iPSCs have immunogenic features and the immunogenicity of therapeutically of these cells should be evaluated before any clinical application.

    Article  CAS  PubMed  Google Scholar 

  54. Deng L, Kong X, Liu G, Li C, Chen H, Hong Z, et al. Transplantation of adipose-derived mesenchymal stem cells efficiently rescues thioacetamide-induced acute liver failure in mice. Transplant Proc. 2016;48:2208–15.

  55. Shi D, Zhang J, Zhou Q, Xin J, Jiang J, Jiang L, et al. Quantitative evaluation of human bone mesenchymal stem cells rescuing fulminant hepatic failure in pigs. Gut. 2017;66:955–64.

  56. LIU T, MU H, SHEN Z, SONG Z, CHEN X, WANG Y. Autologous adipose tissue-derived mesenchymal stem cells are involved in rat liver regeneration following repeat partial hepatectomy. Mol Med Rep. 2016;13:2053–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Tautenhahn H-M, Brückner S, Uder C, Erler S, Hempel M, von Bergen M, et al. Mesenchymal stem cells correct haemodynamic dysfunction associated with liver injury after extended resection in a pig model. Sci Rep. 2017;7:2617.

  58. Liao N, et al. Adipose tissue-derived stem cells promote the reversion of non-alcoholic fatty liver disease: an in vivo study. Int J Mol Med. 2016;37:1389–96.

    Article  CAS  PubMed  Google Scholar 

  59. Tautenhahn H-M, Brückner S, Baumann S, Winkler S, Otto W, von Bergen M, et al. Attenuation of postoperative acute liver failure by mesenchymal stem cell treatment due to metabolic implications. Ann Surg. 2016;263:546–56.

  60. Huang B, Cheng X, Wang H, Huang W, la Ga hu Z, Wang D, et al. Mesenchymal stem cells and their secreted molecules predominantly ameliorate fulminant hepatic failure and chronic liver fibrosis in mice respectively. J Transl Med. 2016;14:45.

  61. Gilsanz C, Aller MA, Fuentes-Julian S, Prieto I, Blázquez-Martinez A, Argudo S, et al. Adipose-derived mesenchymal stem cells slow disease progression of acute-on-chronic liver failure. Biomed Pharmacother. 2017;91:776–87.

  62. Kaur S, Siddiqui H, Bhat MH. Hepatic progenitor cells in action: liver regeneration or fibrosis? Am J Pathol. 2015;185:2342–50.

    Article  CAS  PubMed  Google Scholar 

  63. Asawa S, Saito T, Satoh A, Ohtake K, Tsuchiya T, Okada H, et al. Participation of bone marrow cells in biliary fibrosis after bile duct ligation. J Gastroenterol Hepatol Aust. 2007;22:2001–8.

  64. Jang YO, Jun BG, Baik SK, Kim MY, Kwon SO. Inhibition of hepatic stellate cells by bone marrow-derived mesenchymal stem cells in hepatic fibrosis. Clin Mol Hepatol. 2015;21:141.

    Article  PubMed  PubMed Central  Google Scholar 

  65. Yu F, Ji S, Su L, Wan L, Zhang S, Dai C, et al. Adipose-derived mesenchymal stem cells inhibit activation of hepatic stellate cells in vitro and ameliorate rat liver fibrosis in vivo. J Formos Med Assoc. 2015;114:130–8.

  66. Zhang L-T, et al. Bone marrow-derived mesenchymal stem cells inhibit the proliferation of hepatic stellate cells by inhibiting the transforming growth factor β pathway. Mol Med Rep. 2015;12:7227–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Tang W-P, Akahoshi T, Piao JS, Narahara S, Murata M, Kawano T, et al. Basic fibroblast growth factor-treated adipose tissue-derived mesenchymal stem cell infusion to ameliorate liver cirrhosis via paracrine hepatocyte growth factor: ADSCs relieve liver cirrhosis. J Gastroenterol Hepatol. 2015;30:1065–74.

  68. Nagamoto Y, Takayama K, Ohashi K, Okamoto R, Sakurai F, Tachibana M, et al. Transplantation of a human iPSC-derived hepatocyte sheet increases survival in mice with acute liver failure. J Hepatol. 2016;64:1068–75.

  69. Kubo K, Ohnishi S, Hosono H, Fukai M, Kameya A, Higashi R, et al. Human amnion-derived mesenchymal stem cell transplantation ameliorates liver fibrosis in rats. Transplant Direct. 2015;1:1–9.

  70. Liang H, Huang K, Su T, Li Z, Hu S, Dinh PU, et al. Mesenchymal stem cell/red blood cell-inspired nanoparticle therapy in mice with carbon tetrachloride-induced acute liver failure. ACS Nano. 2018;12:6536–44.

  71. Thomson JA, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998;282:1145–7.

    Article  CAS  PubMed  Google Scholar 

  72. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131:861–72.

  73. Agarwal S, Holton KL, Lanza R. Efficient differentiation of functional hepatocytes from human embryonic stem cells. Stem Cells Dayt Ohio. 2008;26:1117–27.

  74. Dhawan A, Puppi J, Hughes RD, Mitry RR. Human hepatocyte transplantation: current experience and future challenges. Nat Rev Gastroenterol Hepatol. 2010;7:288–98.

    Article  PubMed  Google Scholar 

  75. Babaei A, Katoonizadeh A, Ranjbar A, Naderi M, Ahmadbeigi N. Directly injected native bone-marrow stem cells cannot incorporate into acetaminophen-induced liver injury. Biologicals. 2018;52:55–8.

    Article  PubMed  Google Scholar 

  76. Ribes-Koninckx C, Ibars EP, Agrasot MÁC, Bonora-Centelles A, Miquel BP, Carbó JJV, et al. Clinical outcome of hepatocyte transplantation in four pediatric patients with inherited metabolic diseases. Cell Transplant. 2012;21:2267–82.

  77. Mooney DJ, Vandenburgh H. Cell delivery mechanisms for tissue repair. Cell Stem Cell. 2008;2:205–13.

    Article  CAS  PubMed  Google Scholar 

  78. Nicolas CT, Hickey RD, Chen HS, Mao SA, Lopera Higuita M, Wang Y, et al. Concise review: liver regenerative medicine: from hepatocyte transplantation to bioartificial livers and bioengineered grafts: liver regenerative medicine. Stem Cells. 2017;35:42–50.

  79. Smets F, Najimi M, Sokal EM. Cell transplantation in the treatment of liver diseases. Pediatr Transplant. 2008;12:6–13.

    Article  CAS  PubMed  Google Scholar 

  80. Song W, Lu YC, Frankel AS, An D, Schwartz RE, Ma M. Engraftment of human induced pluripotent stem cell-derived hepatocytes in immunocompetent mice via 3D co-aggregation and encapsulation. Sci Rep. 2015;5.

Download references

Author information

Authors and Affiliations

  1. Center for Engineering in Medicine, Massachusetts General Hospital, Shriners Hospitals for Children, Harvard Medical School, 51 Blossom Street, Boston, MA, 02114, USA

    Aylin Acun, Ruben Oganesyan & Basak E. Uygun

Authors
  1. Aylin Acun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ruben Oganesyan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Basak E. Uygun
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Basak E. Uygun.

Ethics declarations

Conflict of Interest

Basak Uygun reports grants from National Institutes of Health (R01DK084053) and has a financial interest in Organ Solutions, LLC (reviewed and arranged by MGH and Partners HealthCare in accordance with their conflict of interest policies).

Aylin Acun and Ruben Oganesyan declare no conflict of interest.

Human and Animal Rights and Informed Consent

This article does not contain any studies with human or animal subjects performed by any of the authors.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

This article is part of the Topical Collection on Cellular Transplants

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Acun, A., Oganesyan, R. & Uygun, B.E. Liver Bioengineering: Promise, Pitfalls, and Hurdles to Overcome. Curr Transpl Rep 6, 119–126 (2019). https://doi.org/10.1007/s40472-019-00236-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40472-019-00236-3

Keywords

Search

Navigation