Background:

Organ bioengineering has the potential to completely eliminate the donor organ shortage. Here, we examine the feasibility of a decellularized liver construct to support the growth of multilineage allogenic functional cells.

Methods:

Multilineage neonatal rat liver cell slurry was introduced into decellularized adult rat livers scaffolds, via the bile duct. Recellularized grafts were perfused with cell growth medium through the portal vein for 7 days. Concurrently, the same cell slurry was incubated on culture dishes. Albumin levels were measured from grafts perfusates and cell culture media. Immunofluorescent assays were used to detect cholangiocytes, hepatocytes and endothelial cells in the recellularized grafts using anti-CK-7, anti-hepatocyte antigen and anti-CD34, respectively.

Results:

The colocalization of cholangiocytes, hepatocytes and endothelial cells were verified by cell markers positivity. The perfusate from three grafts showed increasing albumin concentration over seven days. Albumin production was higher in graft perfusate relative to that in culture dishes.Conclusion:

Biological scaffolds supported the growth of three different liver cell lines. Recellularized grafts were metabolically functional as demonstrated by albumin production,representing a promising model for transplantable biological construct.(A)Native rat liver stained for cholangiocytes(green),(B)Endothelial cells(red). (C)Recellularized grafts with neonatal rat liver cell slurry stained for cholangiocytes(green)and hepatocytes(pink), (D)Endothelial cells(Red) and (E)Merged image of(C)&(D)Albumin levels in perfusates of liver constructs(n=3) and cell slurry in culture dishes.

CITATION INFORMATION: Hassanein W, Werdesheim A, Cimeno A, Buckingham B, Khalifeh A, Uluer M, Harrison J, Parsell D, Drachenberg C, Barth R, LaMattina J. Decellularized Rat Liver Scaffolds Support Survival and Metabolic Function of Multilineage Allogenic Cells. Am J Transplant. 2017;17 (suppl 3).

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