Preparation of Decellularized Liver Scaffolds and Recellularized Liver Grafts

  • Yibin Chen
  • Sharon Geerts
  • Maria Jaramillo
  • Basak E. Uygun
Part of the Methods in Molecular Biology book series (MIMB, volume 1577)


Severe liver disease is the 12th leading cause of death in the USA, with organ transplantation often being the only viable option for treatment. However, due to the shortage of viable donor livers, it is estimated that over 1200 patients died in 2015 while waiting for liver transplantation. This highlights the need for alternative sources of viable organs. In this study, we describe a method that provides the groundwork for the development of functional liver grafts. The approach described here is for removal of cells from intact livers and subsequently repopulating them with functional liver cells. Briefly, rat livers are harvested and subjected to a series of perfusion decellularization steps using an anionic detergent such that an intact decellularized liver matrix (DLM) scaffold with preserved vascular architecture is obtained. Further, we describe methods to recellularize DLM scaffolds with adult primary hepatocytes, creating a liver graft that exhibits hepatic functions in vitro.


Decellularization Liver tissue engineering Recellularization Whole liver scaffold 


  1. 1.
    Badylak SF, Taylor D, Uygun K (2011) Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds. Annu Rev Biomed Eng 13:27–53CrossRefGoogle Scholar
  2. 2.
    Gilbert TW, Sellaro TL, Badylak SF (2006) Decellularization of tissues and organs. Biomaterials 27:3675–3683PubMedGoogle Scholar
  3. 3.
    Macchiarini P, Jungebluth P, Go T et al (2008) Clinical transplantation of a tissue-engineered airway. Lancet 372:2023–2030CrossRefGoogle Scholar
  4. 4.
    Ott HC, Matthiesen TS, Goh S-K et al (2008) Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med 14:213–221CrossRefGoogle Scholar
  5. 5.
    Petersen TH, Calle EA, Zhao L et al (2010) Tissue-engineered lungs for in vivo implantation. Science 329:538–541CrossRefGoogle Scholar
  6. 6.
    Ott HC, Clippinger B, Conrad C et al (2010) Regeneration and orthotopic transplantation of a bioartificial lung. Nat Med 16:927–933CrossRefGoogle Scholar
  7. 7.
    Sullivan DC, Mirmalek-Sani S-H, Deegan DB et al (2012) Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system. Biomaterials 33:7756–7764CrossRefGoogle Scholar
  8. 8.
    Bolland F, Korossis S, Wilshaw S-P et al (2007) Development and characterisation of a full-thickness acellular porcine bladder matrix for tissue engineering. Biomaterials 28:1061–1070CrossRefGoogle Scholar
  9. 9.
    Uygun BE, Soto-Gutierrez A, Yagi H et al (2010) Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med 16:814–820CrossRefGoogle Scholar
  10. 10.
    Uygun BE, Izamis M-L, Jaramillo M et al (2016) Discarded livers find a new life: engineered liver grafts using hepatocytes recovered from marginal livers. Artif Organs 13:27Google Scholar
  11. 11.
    Dunn JC, Tompkins RG, Yarmush ML (1991) Long-term in vitro function of adult hepatocytes in a collagen sandwich configuration. Biotechnol Prog 7:237–245CrossRefGoogle Scholar
  12. 12.
    Farndale RW, Buttle DJ, Barrett AJ (1986) Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 883:173–177CrossRefGoogle Scholar
  13. 13.
    Geerts S, Ozer S, Jaramillo M et al (2016) Nondestructive methods for monitoring cell removal during rat liver decellularization. Tissue Eng Part C Meth 22:671–678CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Yibin Chen
    • 1
    • 2
  • Sharon Geerts
    • 1
    • 2
  • Maria Jaramillo
    • 1
    • 2
  • Basak E. Uygun
    • 1
    • 2
  1. 1.Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical SchoolBostonUSA
  2. 2.Shriners Hospitals for Children in BostonBostonUSA

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