Background:

Recent years have seen a proliferation of methods leading to successful organ decellularization. Nonetheless, the ability to reproducibly generate a robust recellularized construct remains challenging. Here we demonstrate the ability of rat liver scaffolds to support the growth of a variety of human and rat cells.

Methods:

Biochamber systems were used to perfuse adult rat livers with 0.1% SDS for 24 hours yielding decellularized liver scaffolds, preserving the extracellular matrix. Initial studies were performed using a human tumor cell line (hepG2 through the bile duct) due to ease of cell culture. A second series utilized human tumor cells co-cultured with human umbilical vein endothelial cells (HUVECs, introduced via the portal vein). The third series rat neonatal cell slurry was seeded on the scaffold providing all cell types found in the liver. Neonatal hepatocytes were isolated from 3 day old neonatal rats using a modified two step isolation technique and perfused onto liver scaffolds via the bile duct. Biochambers were used to circulate oxygenated hepatocyte basal medium via the portal vein at 37C for 5 days.

Results:

Human HepG2 cells grew readily on the scaffold (n=20). HepG2 cells co-cultured with HUVECs demonstrated viable endothelial lining with concurrent hepatocyte growth (n=10, Figure 1). In the series of neonatal cell slurry infusion (n=9), obvious foci of neonatal hepatocytes were observed to repopulate the parenchyma of the scaffold (Figure 2).

Conclusion:

In an appropriately selected media rat liver scaffolds support human cell ingrowth, engraftment and survival of a neonatal rat liver cell slurry. Further work characterizing the function and long-term viability of the construct is underway.

CITATION INFORMATION: Hassanein W, Dhru U, Woodall J, Uluer M, Cimeno A, Parsell D, Rivera C, Klepfer S, Drachenberg C, Barth R, LaMattina J. Decellularized Rat Liver Scaffolds Support Multilineage Xenogenic and Heterogenic Cell Growth. Am J Transplant. 2016;16 (suppl 3).

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