TY - JOUR
T1 - A whole-organ regenerative medicine approach for liver replacement
AU - Soto-Gutierrez, Alejandro
AU - Zhang, Li
AU - Medberry, Chris
AU - Fukumitsu, Ken
AU - Faulk, Denver
AU - Jiang, Hongbin
AU - Reing, Janet
AU - Gramignoli, Roberto
AU - Komori, Junji
AU - Ross, Mark
AU - Nagaya, Masaki
AU - Lagasse, Eric
AU - Stolz, Donna
AU - Strom, Stephen C.
AU - Fox, Ira J.
AU - Badylak, Stephen F.
PY - 2011/6/1
Y1 - 2011/6/1
N2 - Background & Aims: The therapy of choice for end-stage liver disease is whole-organ liver transplantation, but this option is limited by a shortage of donor organs. Cell-based therapies and hepatic tissue engineering have been considered as alternatives to liver transplantation, but neither has proven effective to date. A regenerative medicine approach for liver replacement has recently been described that includes the use of a three-dimensional organ scaffold prepared by decellularization of xenogeneic liver. The present study investigates a new, minimally disruptive method for whole-organ liver decellularization and three different cell reseeding strategies to engineer functional liver tissue. Methods: A combination of enzymatic, detergent, and mechanical methods are used to remove all cells from isolated rat livers. Whole-organ perfusion is used in a customized organ chamber and the decellularized livers are examined by morphologic, biochemical, and immunolabeling techniques for preservation of the native matrix architecture and composition. Three different methods for hepatocyte seeding of the resultant three-dimensional liver scaffolds are evaluated to maximize cell survival and function: (1) direct parenchymal injection, (2) multistep infusion, or (3) continuous perfusion. Results: The decellularization process preserves the three-dimensional macrostructure, the ultrastructure, the composition of the extracellular matrix components, the native microvascular network of the liver, and the bile drainage system, and up to 50% of growth factor content. The three-dimensional liver matrix reseeded with the multistep infusion of hepatocytes generated ∼90% of cell engraftment and supported liver-specific functional capacities of the engrafted cells, including albumin production, urea metabolism, and cytochrome P450 induction. Conclusions: Whole-organ liver decellularization is possible with maintenance of structure and composition suitable to support functional hepatocytes.
AB - Background & Aims: The therapy of choice for end-stage liver disease is whole-organ liver transplantation, but this option is limited by a shortage of donor organs. Cell-based therapies and hepatic tissue engineering have been considered as alternatives to liver transplantation, but neither has proven effective to date. A regenerative medicine approach for liver replacement has recently been described that includes the use of a three-dimensional organ scaffold prepared by decellularization of xenogeneic liver. The present study investigates a new, minimally disruptive method for whole-organ liver decellularization and three different cell reseeding strategies to engineer functional liver tissue. Methods: A combination of enzymatic, detergent, and mechanical methods are used to remove all cells from isolated rat livers. Whole-organ perfusion is used in a customized organ chamber and the decellularized livers are examined by morphologic, biochemical, and immunolabeling techniques for preservation of the native matrix architecture and composition. Three different methods for hepatocyte seeding of the resultant three-dimensional liver scaffolds are evaluated to maximize cell survival and function: (1) direct parenchymal injection, (2) multistep infusion, or (3) continuous perfusion. Results: The decellularization process preserves the three-dimensional macrostructure, the ultrastructure, the composition of the extracellular matrix components, the native microvascular network of the liver, and the bile drainage system, and up to 50% of growth factor content. The three-dimensional liver matrix reseeded with the multistep infusion of hepatocytes generated ∼90% of cell engraftment and supported liver-specific functional capacities of the engrafted cells, including albumin production, urea metabolism, and cytochrome P450 induction. Conclusions: Whole-organ liver decellularization is possible with maintenance of structure and composition suitable to support functional hepatocytes.
UR - http://www.scopus.com/inward/record.url?scp=79958288464&partnerID=8YFLogxK
U2 - 10.1089/ten.tec.2010.0698
DO - 10.1089/ten.tec.2010.0698
M3 - Article
C2 - 21375407
AN - SCOPUS:79958288464
SN - 1937-3384
VL - 17
SP - 677
EP - 686
JO - Tissue Engineering - Part C: Methods
JF - Tissue Engineering - Part C: Methods
IS - 6
ER -