Production and characterization of acellular human liver matrix: Potential auxiliary liver graft

<p>Abdul Hakeem, Wilshaw S P, Ingham E and Prasad K R</p>

Production and characterization of acellular human liver matrix: Potential auxiliary liver graft

Abdul Hakeem, Wilshaw S P, Ingham E and Prasad K R

University of Leeds, UK

: Biomater Med Appl

Abstract

Shortage of donor organs combined with ever increasing liver disease has directed attention towards growing implantable liver tissue in the laboratory setting. Recent developments in decellularization and recellularization techniques have opened exciting facets for liver whole organ reengineering. The aim of this study was to develop a decellularization protocol for human liver, assess the structural and biochemical integrity and assess the biocompatibility of the acellular matrix. Discarded human livers were split and the left lateral lobes with a mean weight of 399±162 g were used. Perfusion decellularization was carried out through the hepatic artery and portal vein. Hypotonic buffer, 0.1% (w/v) sodium dodecyl sulphate, hypertonic buffer and nuclease solutions were used to remove cells plus nuclear material from the liver tissue. Following several iterations of a previously patented Leeds University decellularization protocol, greater than 91.0% DNA was removed from all areas of liver tissue. Histological analysis demonstrated lack of cells and maintenance of the portal triad histio-architecture. Immunohistochemistry demonstrated positive staining for key extracellular proteins such as collagen type I and III, fibronectin and laminin. Scanning electron microscopy showed an intact extracellular matrix. Biochemical assessment confirmed significant reduction in glycosaminoglycan content, but there was increase in collagen content, the latter likely due to extraction by dry weight of other soluble components in the matrix. The acellular tissues and extracts were not cytotoxic to either murine 3T3 or baby hamster kidney cells. Preliminary recellularization work showed that the matrix is biocompatible to primary cryopreserved human hepatocytes seeded at 30,000 cells per m2 density. The study has developed a suitable protocol for decellularization of the left lateral lobe of human liver, without adversely affecting the extracellular matrix structure. The biocompatible acellular scaffold has the potential to be decellularized with hepatocytes, with a view to engineering a transplantable auxiliary liver graft.