studies

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 381A
hepatic injury, increase macrophage recruitment, and increase
hepatic expression of CXCL1 following IRI. The dramatic reduction
in tissue injury by MSC-EV support the therapeutic use of
MSC-EV to reduce hepatic IRI following hepatic surgery or in
hepatic transplantation.
Disclosures:
The following authors have nothing to disclose: Hiroaki Haga, David D. Lee,
Sarah Nix, Irene K. Yan, Tushar Patel
336
Liver Fabrication Using Decellularized Whole-Organ
Scaffold for Transplantable Graft in large animal model
Hiroshi Yagi, Kenta Inomata, kazuki tajima, Taizo Hibi, Yuta Abe,
Minoru Kitago, Masahiro Shinoda, Osamu Itano, Yuko Kitagawa;
Surgery, Keio University School of Medicine, Shinjuku-ku, Japan
Background: Whole-organ scaffold generated by tissue decellularization
technology can be one of the solutions to fabricate
transplantable organ graft, which contains functional cell
types including progenitor cells. However, the feasibility of this
technology for human-scale application, and the biological
alterations of the scaffold after implantation are still unclear.
Methods: To determine if the decellularization methodology
could be applied in large animal model and it could be sufficiently
recellularized with primary cells as well as human iPS
cell derived hepatic progenitor cells, the decellularized scaffold
was first generated using SDS, TrytonX-100 and CHAPS.
Next, endothelial cells and primary porcine hepatocytes were
infused into the scaffold under the monitoring of intravascular
pressure. Finally, the fabricated liver was implanted into the
porcine body and evaluated by angiography after vascular
anastomoses. Histological study was performed to evaluate cell
infiltration, degree of coagulation and adhesion of the scaffold.
In addition, the liver scaffold containing human iPS cell derived
liver progenitor cells was transplanted into porcine body under
immune-suppression. Results: The endothelial cells could cover
most of the vessel lumen of the scaffold and the primary hepatocytes
were well distributed into the parenchymal space. The
fabricated liver could be successfully transplanted into porcine
body connecting with portal vein and IVC. The graft was well
perfused and preserved in the porcine abdominal cavity without
bleeding or absorption. Interestingly, histological analysis
of transplanted scaffold with human iPS derived progenitor
cells revealed liver specific architecture with ALB positive cells
in parenchymal space. Conclusions: We could scale-up and
optimize the liver fabrication system to apply not only primary
cells but also human iPS derived cells into clinically feasible
model.
Disclosures:
The following authors have nothing to disclose: Hiroshi Yagi, Kenta Inomata,
kazuki tajima, Taizo Hibi, Yuta Abe, Minoru Kitago, Masahiro Shinoda, Osamu
Itano, Yuko Kitagawa
337
Three-dimensional bioactive human liver acellular scaffold
with preserved architecture and biomechanical
properties
Giuseppe Mazza 1 , Walid Al-Akkad 1 , Lisa Longato 1 , Andrea
Telese 1 , Andrew R. Hall 1 , Luca Urbani 2 , Benjamin Robinson 3 ,
Giusi Marrone 1 , Oliver Willacy 1 , Luca Frenguelli 1 , Marco Curti 1 ,
Massimo Malago 1 , Tu Vinh Luong 1 , Kevin Moore 1 , Armando E.
Del Rio Hernandez 3 , Paolo De Coppi 2 , Krista Rombouts 1 , Massimo
Pinzani 1 ; 1 Institute for Liver and Digestive Helath, UCL, London,
United Kingdom; 2 Institute for Child Health, UCL, London, United
Kingdom; 3 Imperial College, London, United Kingdom
Background: Human tissue engineering combines cells and
extracellular matrix (ECM) saffolds for the development
of 3D-structure in order to regenerate tissues/organs and
to recapitulate disease in vitro. The key challenge in tissue
engineering is the development of biomaterials that mimic
the complexity of human tissue. Techniques for tissue decellularization
have been introduced in order to obtain natural
acellular scaffold. Optimal scaffolds should be characterised
by preserved ECM integrity, bioactivity and three-dimensional
organisation. Aim: The aim of this study was to develop a rapid
protocol for the decellularisation of small samples of healthy
human liver and demonstrate preservation of ECM composition,
3D-architecture, bioactivity and biomechanical properties.
In addition, repopulation with cultured human liver cell lines,
namely hepatoblastoma (HepG2), hepatic stellate (LX2) cells,
and human umbilical vein endothelial cells (HUVEC). Methods:
Decellularisation of healthy human liver cubes (i.e. 125mm 3 )
was completed within 3 hours of agitation in solutions with
detergents and enzymes. The decellularisation efficiency was
determined by immunohistochemistry for ECM components and
residual DNA, scanning electron and second harmonic generation
microscopy, proteomic, raman spectroscopy, atomic force
microscopy and chorioallantoic membrane assay. RESULTS:
This innovative protocol resulted in liver cube scaffolds with
a preserved 3D structure and ECM composition, while DNA
and cellular residues were successfully removed. Tissue stiffness
was preserved in the decellularized tissues (2KPa) and
the acellular liver maintained the capability to induce neo-angiogenesis
after 7 days. Interestingly, HUVEC repopulated the
decellularised vessels within the liver scaffold expressing functional
marker such as FVIII. Acellular human liver repopulated
with LX2 and HepG2 cells showed remarkable difference in
gene and protein expression when compared with 2D-system.
COL1A1 gene was less expressed in LX2 cells when cultured
in 3D while TGFB1 and LOX were increased. Notably, albumin
mRNA expression was strongly upregulated in HepG2
cells after 14 days in 3D-culture. CONCLUSION: This is the first
report describing a protocol for fast human liver tissue cubes
decellularization. The decellularization protocol maintained the
natural 3D structure and ECM composition and organisation of
human liver tissue. This is a key advance in the development
of 3D technologies for the study of liver disease as well as for
the development of 3D-carrier for hepatocyte transplantation.
Disclosures:
Kevin Moore - Advisory Committees or Review Panels: Servier
Massimo Pinzani - Advisory Committees or Review Panels: Intercept Pharmaceutical,
Silence Therapeutic, Abbot; Consulting: UCB; Speaking and Teaching:
Gilead, BMS