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Simple and quick method for whole-liver decellularization: a novel in vitro three-dimensional bioengineering tool?

  • Toxicokinetics and Metabolism
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Abstract

Proof of principle of organ reengineering through the development of a transplantable recellularized liver graft was published recently. As the decellularization time of the rat liver took 72 h, loss of some key matrix proteins seemed inevitable. Here, we describe the development of a three-dimensional naturally derived liver scaffold with an intact microvascular system that is capable of withstanding fluid flows in the three hepatic circular systems and that is obtained within 60 min. For this purpose, whole rat livers were sequentially perfused with a selection of mild tensioactive substances to remove the cellular components while preserving the major extracellular matrix proteins, including laminin, collagen I, collagen IV, and fibronectin. In addition, we could show the presence of extracellular matrix–bound growth factor islets, important for cell engraftment, migration, proliferation, and differentiation. This easy to prepare scaffold could represent a remarkable tool in the bioengineering of complex three-dimensional in vitro systems for advanced preclinical drug development.

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Abbreviations

DLM:

Decellularized liver matrix

ECM:

Extracellular matrix

KHB:

Krebs-Henseleit buffer

PBS:

Phosphate-buffered saline

SEM:

Scanning electron microscopy

SDS:

Sodium dodecyl sulfate

3D:

Three-dimensional

VEGF:

Vascular endothelial growth factor

References

  • Badylak SF (2007) The extracellular matrix as a biologic scaffold material. Biomaterials 28(25):3587–3593

    Article  PubMed  CAS  Google Scholar 

  • De Spiegelaere W, Casteleyn C, Van Den Broeck W, Simoens P (2008) Electron microscopic study of the porcine choroid plexus epithelium. Anat Histol Embryol 37:458–463

    Article  PubMed  Google Scholar 

  • De Spiegelaere W, Cornillie P, Casteleyn C, Burvenich C, Van den Broeck W (2010) Detection of hypoxia inducible factors and angiogenic growth factors during foetal endochondral and intramembranous ossification. Anat Histol Embryol 39:376–384

    Article  PubMed  Google Scholar 

  • Gómez-Lechón MJ, Lahoz A, Gombau L, Castell JV, Donato MT (2010) In vitro evaluation of potential hepatotoxicity induced by drugs. Curr Pharm Des 16(17):1963–1977

    Article  PubMed  Google Scholar 

  • Guguen-Guillouzo C, Guillouzo A (2010) General review on in vitro hepatocyte models and their applications. Methods Mol Biol 640:1–40

    Article  PubMed  CAS  Google Scholar 

  • Henkens T, Vanhaecke T, Papeleu P, Elaut G, Vinken M, Snykers S, Rogiers V (2006) Rat hepatocyte cultures: conventional monolayer and cocultures with rat liver epithelial cells. Methods Mol Biol 320:239–246

    PubMed  Google Scholar 

  • Järveläinen H, Sainio A, Koulu M, Wight TN, Penttinen R (2009) Extracellular matrix molecules: potential targets in pharmacotherapy. Pharmacol Rev 61(2):198–223

    Article  PubMed  Google Scholar 

  • Papeleu P, Vanhaecke T, Henkens T, Elaut G, Vinken M, Snykers S, Rogiers V (2006) Isolation of rat hepatocytes. Methods Mol Biol 320: 229–237

    PubMed  Google Scholar 

  • Park JE, Keller GA, Ferrara N (1993) The vascular endothelial growth factor (VEGF) isoforms: differential deposition into the subepithelial extracellular matrix and bioactivity of extracellular matrix-bound VEGF. Mol Biol Cell 4(12):1317–1326

    PubMed  CAS  Google Scholar 

  • Uygun BE, Soto-Gutierrez A, Yagi H, Izamis ML, Guzzardi MA, Shulman C, Milwid J, Kobayashi N, Tilles A, Berthiaume F, Hertl M, Nahmias Y, Yarmush ML, Uygun K (2010) Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med 16(7):814–820

    Article  PubMed  CAS  Google Scholar 

  • Vinken M, Elaut G, Henkens T, Papeleu P, Snykers S, Vanhaecke T, Rogiers V (2006) Rat hepatocyte cultures: collagen gel sandwich and immobilization cultures. Methods Mol Biol 320:247–254

    PubMed  CAS  Google Scholar 

  • Wang S, Nagrath D, Chen PC, Berthiaume F, Yarmush ML (2008) Three-dimensional primary hepatocyte culture in synthetic self-assembling peptide hydrogel. Tissue Eng 14(2):227–236

    Article  CAS  Google Scholar 

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Acknowledgments

The authors thank Tatyana Doktorova, Jurgen De Craene, and Godelieve De Pauw for their excellent technical assistance.

Financial support

Joery De Kock is a doctoral research fellow of the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen). The research leading to these results has also received funding from the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n°20161 (ESNATS) and from ISRIB (Brustem) and BELSPO (IAP).

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The authors state that no conflict of interest is applicable to the presented work.

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Correspondence to Joery De Kock.

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De Kock, J., Ceelen, L., De Spiegelaere, W. et al. Simple and quick method for whole-liver decellularization: a novel in vitro three-dimensional bioengineering tool?. Arch Toxicol 85, 607–612 (2011). https://doi.org/10.1007/s00204-011-0706-1

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  • DOI: https://doi.org/10.1007/s00204-011-0706-1

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