title: Recellularization via the bile duct supports functional allogenic and xenogenic cell growth on a decellularized rat liver scaffold.
abstract: Recent years have seen a proliferation of methods leading to successful organ decellularization. In this experiment we examine the feasibility of a decellularized liver construct to support growth of functional multilineage cells. Bio-chamber systems were used to perfuse adult rat livers with 0.1% SDS for 24 hours yielding decellularized liver scaffolds. Initially, we recellularized liver scaffolds using a human tumor cell line (HepG2, introduced via the bile duct). Subsequent studies were performed using either human tumor cells co-cultured with human umbilical vein endothelial cells (HUVECs, introduced via the portal vein) or rat neonatal cell slurry (introduced via the bile duct). Bio-chambers were used to circulate oxygenated growth medium via the portal vein at 37C for 5-7 days. Human HepG2 cells grew readily on the scaffold (n = 20). HepG2 cells co-cultured with HUVECs demonstrated viable human endothelial lining with concurrent hepatocyte growth (n = 10). In the series of neonatal cell slurry infusion (n = 10), distinct foci of neonatal hepatocytes were observed to repopulate the parenchyma of the scaffold. The presence of cholangiocytes was verified by CK-7 positivity. Quantitative albumin measurement from the grafts showed increasing albumin levels after seven days of perfusion. Graft albumin production was higher than that observed in traditional cell culture. This data shows that rat liver scaffolds support human cell ingrowth. The scaffold likewise supported the engraftment and survival of neonatal rat liver cell slurry. Recellularization of liver scaffolds thus presents a promising model for functional liver engineering.
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Wessam
authors.initials: W
authors.lastname: Hassanein
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Mehmet C
authors.initials: MC
authors.lastname: Uluer
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: John
authors.initials: J
authors.lastname: Langford
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Jhade D
authors.initials: JD
authors.lastname: Woodall
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Arielle
authors.initials: A
authors.lastname: Cimeno
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Urmil
authors.initials: U
authors.lastname: Dhru
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Avraham
authors.initials: A
authors.lastname: Werdesheim
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Harrison
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Carlos
authors.initials: C
authors.lastname: Rivera-Pratt
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: Klepfer
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Ali
authors.initials: A
authors.lastname: Khalifeh
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Bryan
authors.initials: B
authors.lastname: Buckingham
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Philip S
authors.initials: PS
authors.lastname: Brazio
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Dawn
authors.initials: D
authors.lastname: Parsell
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Charlie
authors.initials: C
authors.lastname: Klassen
authors.affiliation: b University of Maryland School of Medicine , Department of Pathology , Baltimore , MD , USA.
authors.forename: Cinthia
authors.initials: C
authors.lastname: Drachenberg
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: Rolf N
authors.initials: RN
authors.lastname: Barth
authors.affiliation: a University of Maryland School of Medicine, Department of Surgery , Division of Transplantation , Baltimore , MD , USA.
authors.forename: John C
authors.initials: JC
authors.lastname: LaMattina
mesh_terms: D000418:Albumins / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001652:Bile Ducts / Q000166:cytology*; D002451:Cell Compartmentation; D019070:Cell Lineage; D049109:Cell Proliferation; D058948:Cell Tracking; D004247:DNA / Q000378:metabolism; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000166:cytology*; D008297:Male; D017208:Rats, Wistar; D015203:Reproducibility of Results; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000418:Albumins; D004247:DNA
keywords: bioengineering; decellularization; liver; organ scaffold; rat; recellularization